CN102516381A - Natural sex storage protein-2 as well as preparation and use for same - Google Patents
Natural sex storage protein-2 as well as preparation and use for same Download PDFInfo
- Publication number
- CN102516381A CN102516381A CN201110426497XA CN201110426497A CN102516381A CN 102516381 A CN102516381 A CN 102516381A CN 201110426497X A CN201110426497X A CN 201110426497XA CN 201110426497 A CN201110426497 A CN 201110426497A CN 102516381 A CN102516381 A CN 102516381A
- Authority
- CN
- China
- Prior art keywords
- storage protein
- supernatant
- sex storage
- natural sex
- phe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 238000003860 storage Methods 0.000 title abstract 4
- 239000006228 supernatant Substances 0.000 claims abstract description 41
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 19
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 19
- 241000255789 Bombyx mori Species 0.000 claims abstract description 18
- 238000004113 cell culture Methods 0.000 claims abstract description 14
- 238000001914 filtration Methods 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims abstract description 8
- 230000031787 nutrient reservoir activity Effects 0.000 claims description 61
- 238000000926 separation method Methods 0.000 claims description 26
- 210000002966 serum Anatomy 0.000 claims description 19
- 238000000108 ultra-filtration Methods 0.000 claims description 19
- 238000000746 purification Methods 0.000 claims description 18
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 12
- 239000012528 membrane Substances 0.000 claims description 11
- 239000003643 water by type Substances 0.000 claims description 9
- 238000004321 preservation Methods 0.000 claims description 8
- 239000012505 Superdex™ Substances 0.000 claims description 7
- 239000012141 concentrate Substances 0.000 claims description 7
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 6
- 238000002203 pretreatment Methods 0.000 claims description 6
- 238000003556 assay Methods 0.000 claims description 5
- 239000006143 cell culture medium Substances 0.000 claims description 5
- 230000008021 deposition Effects 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 4
- 238000012546 transfer Methods 0.000 claims description 4
- 238000003828 vacuum filtration Methods 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 239000002994 raw material Substances 0.000 abstract description 5
- 230000006907 apoptotic process Effects 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 238000011031 large-scale manufacturing process Methods 0.000 abstract 1
- 239000003223 protective agent Substances 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 14
- 238000001962 electrophoresis Methods 0.000 description 9
- 239000000706 filtrate Substances 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 239000001963 growth medium Substances 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- 108090000765 processed proteins & peptides Proteins 0.000 description 6
- 239000012679 serum free medium Substances 0.000 description 6
- 230000008878 coupling Effects 0.000 description 4
- 238000010168 coupling process Methods 0.000 description 4
- 238000005859 coupling reaction Methods 0.000 description 4
- 239000012160 loading buffer Substances 0.000 description 4
- 230000009182 swimming Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 3
- 150000001413 amino acids Chemical group 0.000 description 3
- 230000002424 anti-apoptotic effect Effects 0.000 description 3
- 108010092854 aspartyllysine Proteins 0.000 description 3
- 108010090894 prolylleucine Proteins 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 241000283690 Bos taurus Species 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- QRWPTXLWHHTOCO-DZKIICNBSA-N Glu-Val-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O QRWPTXLWHHTOCO-DZKIICNBSA-N 0.000 description 2
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 2
- SRILZRSXIKRGBF-HRCADAONSA-N Phe-Met-Pro Chemical compound CSCC[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N SRILZRSXIKRGBF-HRCADAONSA-N 0.000 description 2
- GAMLAXHLYGLQBJ-UFYCRDLUSA-N Phe-Val-Tyr Chemical compound N[C@H](C(=O)N[C@H](C(=O)N[C@H](C(=O)O)CC1=CC=C(C=C1)O)C(C)C)CC1=CC=CC=C1 GAMLAXHLYGLQBJ-UFYCRDLUSA-N 0.000 description 2
- AWQGDZBKQTYNMN-IHRRRGAJSA-N Pro-Phe-Asp Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N[C@@H](CC(=O)O)C(=O)O AWQGDZBKQTYNMN-IHRRRGAJSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 239000010975 amethyst Substances 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 230000004637 cellular stress Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 239000010931 gold Substances 0.000 description 2
- 229910052737 gold Inorganic materials 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 231100000957 no side effect Toxicity 0.000 description 2
- 230000000050 nutritive effect Effects 0.000 description 2
- 238000007639 printing Methods 0.000 description 2
- 108010031719 prolyl-serine Proteins 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 108010020532 tyrosyl-proline Proteins 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- FJVAQLJNTSUQPY-CIUDSAMLSA-N Ala-Ala-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN FJVAQLJNTSUQPY-CIUDSAMLSA-N 0.000 description 1
- ZIWWTZWAKYBUOB-CIUDSAMLSA-N Ala-Asp-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O ZIWWTZWAKYBUOB-CIUDSAMLSA-N 0.000 description 1
- DXTYEWAQOXYRHZ-KKXDTOCCSA-N Ala-Phe-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N DXTYEWAQOXYRHZ-KKXDTOCCSA-N 0.000 description 1
- BHTBAVZSZCQZPT-GUBZILKMSA-N Ala-Pro-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N BHTBAVZSZCQZPT-GUBZILKMSA-N 0.000 description 1
- CQJHFKKGZXKZBC-BPNCWPANSA-N Ala-Pro-Tyr Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 CQJHFKKGZXKZBC-BPNCWPANSA-N 0.000 description 1
- IASNWHAGGYTEKX-IUCAKERBSA-N Arg-Arg-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(O)=O IASNWHAGGYTEKX-IUCAKERBSA-N 0.000 description 1
- MFAMTAVAFBPXDC-LPEHRKFASA-N Arg-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O MFAMTAVAFBPXDC-LPEHRKFASA-N 0.000 description 1
- KRQSPVKUISQQFS-FJXKBIBVSA-N Arg-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCN=C(N)N KRQSPVKUISQQFS-FJXKBIBVSA-N 0.000 description 1
- LLUGJARLJCGLAR-CYDGBPFRSA-N Arg-Ile-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N LLUGJARLJCGLAR-CYDGBPFRSA-N 0.000 description 1
- ADPACBMPYWJJCE-FXQIFTODSA-N Arg-Ser-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O ADPACBMPYWJJCE-FXQIFTODSA-N 0.000 description 1
- BECXEHHOZNFFFX-IHRRRGAJSA-N Arg-Ser-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BECXEHHOZNFFFX-IHRRRGAJSA-N 0.000 description 1
- UZSQXCMNUPKLCC-FJXKBIBVSA-N Arg-Thr-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O UZSQXCMNUPKLCC-FJXKBIBVSA-N 0.000 description 1
- XEOXPCNONWHHSW-AVGNSLFASA-N Arg-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N XEOXPCNONWHHSW-AVGNSLFASA-N 0.000 description 1
- PAXHINASXXXILC-SRVKXCTJSA-N Asn-Asp-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N)O PAXHINASXXXILC-SRVKXCTJSA-N 0.000 description 1
- BZMWJLLUAKSIMH-FXQIFTODSA-N Asn-Glu-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O BZMWJLLUAKSIMH-FXQIFTODSA-N 0.000 description 1
- AITGTTNYKAWKDR-CIUDSAMLSA-N Asn-His-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O AITGTTNYKAWKDR-CIUDSAMLSA-N 0.000 description 1
- UYXXMIZGHYKYAT-NHCYSSNCSA-N Asn-His-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC(=O)N)N UYXXMIZGHYKYAT-NHCYSSNCSA-N 0.000 description 1
- XVBDDUPJVQXDSI-PEFMBERDSA-N Asn-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N XVBDDUPJVQXDSI-PEFMBERDSA-N 0.000 description 1
- LTZIRYMWOJHRCH-GUDRVLHUSA-N Asn-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)N)N LTZIRYMWOJHRCH-GUDRVLHUSA-N 0.000 description 1
- NYGILGUOUOXGMJ-YUMQZZPRSA-N Asn-Lys-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O NYGILGUOUOXGMJ-YUMQZZPRSA-N 0.000 description 1
- RAUPFUCUDBQYHE-AVGNSLFASA-N Asn-Phe-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O RAUPFUCUDBQYHE-AVGNSLFASA-N 0.000 description 1
- UYCPJVYQYARFGB-YDHLFZDLSA-N Asn-Phe-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O UYCPJVYQYARFGB-YDHLFZDLSA-N 0.000 description 1
- REQUGIWGOGSOEZ-ZLUOBGJFSA-N Asn-Ser-Asn Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)C(=O)N REQUGIWGOGSOEZ-ZLUOBGJFSA-N 0.000 description 1
- NCXTYSVDWLAQGZ-ZKWXMUAHSA-N Asn-Ser-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O NCXTYSVDWLAQGZ-ZKWXMUAHSA-N 0.000 description 1
- HPASIOLTWSNMFB-OLHMAJIHSA-N Asn-Thr-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O HPASIOLTWSNMFB-OLHMAJIHSA-N 0.000 description 1
- CGYKCTPUGXFPMG-IHPCNDPISA-N Asn-Tyr-Trp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O CGYKCTPUGXFPMG-IHPCNDPISA-N 0.000 description 1
- CBWCQCANJSGUOH-ZKWXMUAHSA-N Asn-Val-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O CBWCQCANJSGUOH-ZKWXMUAHSA-N 0.000 description 1
- JZLFYAAGGYMRIK-BYULHYEWSA-N Asn-Val-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O JZLFYAAGGYMRIK-BYULHYEWSA-N 0.000 description 1
- WQAOZCVOOYUWKG-LSJOCFKGSA-N Asn-Val-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CC(=O)N)N WQAOZCVOOYUWKG-LSJOCFKGSA-N 0.000 description 1
- DBWYWXNMZZYIRY-LPEHRKFASA-N Asp-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(=O)O)N)C(=O)O DBWYWXNMZZYIRY-LPEHRKFASA-N 0.000 description 1
- ICTXFVKYAGQURS-UBHSHLNASA-N Asp-Asn-Trp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O ICTXFVKYAGQURS-UBHSHLNASA-N 0.000 description 1
- RYEWQKQXRJCHIO-SRVKXCTJSA-N Asp-Asn-Tyr Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 RYEWQKQXRJCHIO-SRVKXCTJSA-N 0.000 description 1
- QOVWVLLHMMCFFY-ZLUOBGJFSA-N Asp-Asp-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O QOVWVLLHMMCFFY-ZLUOBGJFSA-N 0.000 description 1
- VAWNQIGQPUOPQW-ACZMJKKPSA-N Asp-Glu-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O VAWNQIGQPUOPQW-ACZMJKKPSA-N 0.000 description 1
- RRKCPMGSRIDLNC-AVGNSLFASA-N Asp-Glu-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O RRKCPMGSRIDLNC-AVGNSLFASA-N 0.000 description 1
- QCVXMEHGFUMKCO-YUMQZZPRSA-N Asp-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(O)=O QCVXMEHGFUMKCO-YUMQZZPRSA-N 0.000 description 1
- KQBVNNAPIURMPD-PEFMBERDSA-N Asp-Ile-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O KQBVNNAPIURMPD-PEFMBERDSA-N 0.000 description 1
- SEMWSADZTMJELF-BYULHYEWSA-N Asp-Ile-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O SEMWSADZTMJELF-BYULHYEWSA-N 0.000 description 1
- IOXWDLNHXZOXQP-FXQIFTODSA-N Asp-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC(=O)O)N IOXWDLNHXZOXQP-FXQIFTODSA-N 0.000 description 1
- JSHWXQIZOCVWIA-ZKWXMUAHSA-N Asp-Ser-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JSHWXQIZOCVWIA-ZKWXMUAHSA-N 0.000 description 1
- CXEFNHOVIIDHFU-IHPCNDPISA-N Asp-Trp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)[C@H](CC(=O)O)N CXEFNHOVIIDHFU-IHPCNDPISA-N 0.000 description 1
- OYSYWMMZGJSQRB-AVGNSLFASA-N Asp-Tyr-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O OYSYWMMZGJSQRB-AVGNSLFASA-N 0.000 description 1
- ALMIMUZAWTUNIO-BZSNNMDCSA-N Asp-Tyr-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ALMIMUZAWTUNIO-BZSNNMDCSA-N 0.000 description 1
- GGBQDSHTXKQSLP-NHCYSSNCSA-N Asp-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)O)N GGBQDSHTXKQSLP-NHCYSSNCSA-N 0.000 description 1
- SBDVXRYCOIEYNV-YUMQZZPRSA-N Cys-His-Gly Chemical compound C1=C(NC=N1)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CS)N SBDVXRYCOIEYNV-YUMQZZPRSA-N 0.000 description 1
- UIKLEGZPIOXFHJ-DLOVCJGASA-N Cys-Phe-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(O)=O UIKLEGZPIOXFHJ-DLOVCJGASA-N 0.000 description 1
- PCRVDEANNSYGTA-IHRRRGAJSA-N Cys-Tyr-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CS)CC1=CC=C(O)C=C1 PCRVDEANNSYGTA-IHRRRGAJSA-N 0.000 description 1
- XOKGKOQWADCLFQ-GARJFASQSA-N Gln-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCC(=O)N)N)C(=O)O XOKGKOQWADCLFQ-GARJFASQSA-N 0.000 description 1
- ODBLJLZVLAWVMS-GUBZILKMSA-N Gln-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)N)N ODBLJLZVLAWVMS-GUBZILKMSA-N 0.000 description 1
- QYTKAVBFRUGYAU-ACZMJKKPSA-N Gln-Asp-Asn Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O QYTKAVBFRUGYAU-ACZMJKKPSA-N 0.000 description 1
- HSHCEAUPUPJPTE-JYJNAYRXSA-N Gln-Leu-Tyr Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N HSHCEAUPUPJPTE-JYJNAYRXSA-N 0.000 description 1
- LURQDGKYBFWWJA-MNXVOIDGSA-N Gln-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(=O)N)N LURQDGKYBFWWJA-MNXVOIDGSA-N 0.000 description 1
- KKCUFHUTMKQQCF-SRVKXCTJSA-N Glu-Arg-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O KKCUFHUTMKQQCF-SRVKXCTJSA-N 0.000 description 1
- LTUVYLVIZHJCOQ-KKUMJFAQSA-N Glu-Arg-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LTUVYLVIZHJCOQ-KKUMJFAQSA-N 0.000 description 1
- QQLBPVKLJBAXBS-FXQIFTODSA-N Glu-Glu-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O QQLBPVKLJBAXBS-FXQIFTODSA-N 0.000 description 1
- YLJHCWNDBKKOEB-IHRRRGAJSA-N Glu-Glu-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O YLJHCWNDBKKOEB-IHRRRGAJSA-N 0.000 description 1
- OAGVHWYIBZMWLA-YFKPBYRVSA-N Glu-Gly-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)NCC(O)=O OAGVHWYIBZMWLA-YFKPBYRVSA-N 0.000 description 1
- KRGZZKWSBGPLKL-IUCAKERBSA-N Glu-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)O)N KRGZZKWSBGPLKL-IUCAKERBSA-N 0.000 description 1
- GRHXUHCFENOCOS-ZPFDUUQYSA-N Glu-Ile-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CCC(=O)O)N GRHXUHCFENOCOS-ZPFDUUQYSA-N 0.000 description 1
- UJMNFCAHLYKWOZ-DCAQKATOSA-N Glu-Lys-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O UJMNFCAHLYKWOZ-DCAQKATOSA-N 0.000 description 1
- QDMVXRNLOPTPIE-WDCWCFNPSA-N Glu-Lys-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QDMVXRNLOPTPIE-WDCWCFNPSA-N 0.000 description 1
- QNJNPKSWAHPYGI-JYJNAYRXSA-N Glu-Phe-Leu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C(O)=O)CC1=CC=CC=C1 QNJNPKSWAHPYGI-JYJNAYRXSA-N 0.000 description 1
- KXTAGESXNQEZKB-DZKIICNBSA-N Glu-Phe-Val Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)CC1=CC=CC=C1 KXTAGESXNQEZKB-DZKIICNBSA-N 0.000 description 1
- CQGBSALYGOXQPE-HTUGSXCWSA-N Glu-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O CQGBSALYGOXQPE-HTUGSXCWSA-N 0.000 description 1
- HJTSRYLPAYGEEC-SIUGBPQLSA-N Glu-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CCC(=O)O)N HJTSRYLPAYGEEC-SIUGBPQLSA-N 0.000 description 1
- FVGOGEGGQLNZGH-DZKIICNBSA-N Glu-Val-Phe Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 FVGOGEGGQLNZGH-DZKIICNBSA-N 0.000 description 1
- WGYHAAXZWPEBDQ-IFFSRLJSSA-N Glu-Val-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WGYHAAXZWPEBDQ-IFFSRLJSSA-N 0.000 description 1
- AQLHORCVPGXDJW-IUCAKERBSA-N Gly-Gln-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)CN AQLHORCVPGXDJW-IUCAKERBSA-N 0.000 description 1
- JUGQPPOVWXSPKJ-RYUDHWBXSA-N Gly-Gln-Phe Chemical compound [H]NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JUGQPPOVWXSPKJ-RYUDHWBXSA-N 0.000 description 1
- BHPQOIPBLYJNAW-NGZCFLSTSA-N Gly-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN BHPQOIPBLYJNAW-NGZCFLSTSA-N 0.000 description 1
- LOEANKRDMMVOGZ-YUMQZZPRSA-N Gly-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)N[C@@H](CC(O)=O)C(O)=O LOEANKRDMMVOGZ-YUMQZZPRSA-N 0.000 description 1
- IEGFSKKANYKBDU-QWHCGFSZSA-N Gly-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)CN)C(=O)O IEGFSKKANYKBDU-QWHCGFSZSA-N 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- LMMPTUVWHCFTOT-GARJFASQSA-N His-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O LMMPTUVWHCFTOT-GARJFASQSA-N 0.000 description 1
- AKEDPWJFQULLPE-IUCAKERBSA-N His-Glu-Gly Chemical compound N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O AKEDPWJFQULLPE-IUCAKERBSA-N 0.000 description 1
- ZSKJIISDJXJQPV-BZSNNMDCSA-N His-Leu-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CN=CN1 ZSKJIISDJXJQPV-BZSNNMDCSA-N 0.000 description 1
- SKOKHBGDXGTDDP-MELADBBJSA-N His-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N SKOKHBGDXGTDDP-MELADBBJSA-N 0.000 description 1
- JUIOPCXACJLRJK-AVGNSLFASA-N His-Lys-Glu Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N JUIOPCXACJLRJK-AVGNSLFASA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- VAXBXNPRXPHGHG-BJDJZHNGSA-N Ile-Ala-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)O)N VAXBXNPRXPHGHG-BJDJZHNGSA-N 0.000 description 1
- UKTUOMWSJPXODT-GUDRVLHUSA-N Ile-Asn-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N UKTUOMWSJPXODT-GUDRVLHUSA-N 0.000 description 1
- HGNUKGZQASSBKQ-PCBIJLKTSA-N Ile-Asp-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N HGNUKGZQASSBKQ-PCBIJLKTSA-N 0.000 description 1
- KYLIZSDYWQQTFM-PEDHHIEDSA-N Ile-Ile-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CCCN=C(N)N KYLIZSDYWQQTFM-PEDHHIEDSA-N 0.000 description 1
- OVDKXUDMKXAZIV-ZPFDUUQYSA-N Ile-Lys-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)N)C(=O)O)N OVDKXUDMKXAZIV-ZPFDUUQYSA-N 0.000 description 1
- HGCNKOLVKRAVHD-UHFFFAOYSA-N L-Met-L-Phe Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 HGCNKOLVKRAVHD-UHFFFAOYSA-N 0.000 description 1
- LJHGALIOHLRRQN-DCAQKATOSA-N Leu-Ala-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N LJHGALIOHLRRQN-DCAQKATOSA-N 0.000 description 1
- RFUBXQQFJFGJFV-GUBZILKMSA-N Leu-Asn-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O RFUBXQQFJFGJFV-GUBZILKMSA-N 0.000 description 1
- ZURHXHNAEJJRNU-CIUDSAMLSA-N Leu-Asp-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ZURHXHNAEJJRNU-CIUDSAMLSA-N 0.000 description 1
- CLVUXCBGKUECIT-HJGDQZAQSA-N Leu-Asp-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CLVUXCBGKUECIT-HJGDQZAQSA-N 0.000 description 1
- APFJUBGRZGMQFF-QWRGUYRKSA-N Leu-Gly-Lys Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN APFJUBGRZGMQFF-QWRGUYRKSA-N 0.000 description 1
- OHZIZVWQXJPBJS-IXOXFDKPSA-N Leu-His-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OHZIZVWQXJPBJS-IXOXFDKPSA-N 0.000 description 1
- HVHRPWQEQHIQJF-AVGNSLFASA-N Leu-Lys-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O HVHRPWQEQHIQJF-AVGNSLFASA-N 0.000 description 1
- RTIRBWJPYJYTLO-MELADBBJSA-N Leu-Lys-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N RTIRBWJPYJYTLO-MELADBBJSA-N 0.000 description 1
- PJWOOBTYQNNRBF-BZSNNMDCSA-N Leu-Phe-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)O)N PJWOOBTYQNNRBF-BZSNNMDCSA-N 0.000 description 1
- FYPWFNKQVVEELI-ULQDDVLXSA-N Leu-Phe-Val Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)CC1=CC=CC=C1 FYPWFNKQVVEELI-ULQDDVLXSA-N 0.000 description 1
- IWMJFLJQHIDZQW-KKUMJFAQSA-N Leu-Ser-Phe Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 IWMJFLJQHIDZQW-KKUMJFAQSA-N 0.000 description 1
- QWWPYKKLXWOITQ-VOAKCMCISA-N Leu-Thr-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(C)C QWWPYKKLXWOITQ-VOAKCMCISA-N 0.000 description 1
- GZRABTMNWJXFMH-UVOCVTCTSA-N Leu-Thr-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZRABTMNWJXFMH-UVOCVTCTSA-N 0.000 description 1
- HGLKOTPFWOMPOB-MEYUZBJRSA-N Leu-Thr-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HGLKOTPFWOMPOB-MEYUZBJRSA-N 0.000 description 1
- RIHIGSWBLHSGLV-CQDKDKBSSA-N Leu-Tyr-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O RIHIGSWBLHSGLV-CQDKDKBSSA-N 0.000 description 1
- WUHBLPVELFTPQK-KKUMJFAQSA-N Leu-Tyr-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O WUHBLPVELFTPQK-KKUMJFAQSA-N 0.000 description 1
- MPOHDJKRBLVGCT-CIUDSAMLSA-N Lys-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N MPOHDJKRBLVGCT-CIUDSAMLSA-N 0.000 description 1
- NFLFJGGKOHYZJF-BJDJZHNGSA-N Lys-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN NFLFJGGKOHYZJF-BJDJZHNGSA-N 0.000 description 1
- IRNSXVOWSXSULE-DCAQKATOSA-N Lys-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN IRNSXVOWSXSULE-DCAQKATOSA-N 0.000 description 1
- YNNPKXBBRZVIRX-IHRRRGAJSA-N Lys-Arg-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O YNNPKXBBRZVIRX-IHRRRGAJSA-N 0.000 description 1
- FACUGMGEFUEBTI-SRVKXCTJSA-N Lys-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCCCN FACUGMGEFUEBTI-SRVKXCTJSA-N 0.000 description 1
- IBQMEXQYZMVIFU-SRVKXCTJSA-N Lys-Asp-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCCN)N IBQMEXQYZMVIFU-SRVKXCTJSA-N 0.000 description 1
- WGCKDDHUFPQSMZ-ZPFDUUQYSA-N Lys-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCCCN WGCKDDHUFPQSMZ-ZPFDUUQYSA-N 0.000 description 1
- IWWMPCPLFXFBAF-SRVKXCTJSA-N Lys-Asp-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O IWWMPCPLFXFBAF-SRVKXCTJSA-N 0.000 description 1
- QIJVAFLRMVBHMU-KKUMJFAQSA-N Lys-Asp-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QIJVAFLRMVBHMU-KKUMJFAQSA-N 0.000 description 1
- IRRZDAIFYHNIIN-JYJNAYRXSA-N Lys-Gln-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IRRZDAIFYHNIIN-JYJNAYRXSA-N 0.000 description 1
- PBIPLDMFHAICIP-DCAQKATOSA-N Lys-Glu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PBIPLDMFHAICIP-DCAQKATOSA-N 0.000 description 1
- VUTWYNQUSJWBHO-BZSNNMDCSA-N Lys-Leu-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VUTWYNQUSJWBHO-BZSNNMDCSA-N 0.000 description 1
- GAHJXEMYXKLZRQ-AJNGGQMLSA-N Lys-Lys-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GAHJXEMYXKLZRQ-AJNGGQMLSA-N 0.000 description 1
- URBJRJKWSUFCKS-AVGNSLFASA-N Lys-Met-Arg Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CCCCN)N URBJRJKWSUFCKS-AVGNSLFASA-N 0.000 description 1
- WKUXWMWQTOYTFI-SRVKXCTJSA-N Lys-Met-Gln Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N WKUXWMWQTOYTFI-SRVKXCTJSA-N 0.000 description 1
- VSTNAUBHKQPVJX-IHRRRGAJSA-N Lys-Met-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O VSTNAUBHKQPVJX-IHRRRGAJSA-N 0.000 description 1
- MTBBHUKKPWKXBT-ULQDDVLXSA-N Lys-Met-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MTBBHUKKPWKXBT-ULQDDVLXSA-N 0.000 description 1
- UDXSLGLHFUBRRM-OEAJRASXSA-N Lys-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CCCCN)N)O UDXSLGLHFUBRRM-OEAJRASXSA-N 0.000 description 1
- HYSVGEAWTGPMOA-IHRRRGAJSA-N Lys-Pro-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O HYSVGEAWTGPMOA-IHRRRGAJSA-N 0.000 description 1
- JHNOXVASMSXSNB-WEDXCCLWSA-N Lys-Thr-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O JHNOXVASMSXSNB-WEDXCCLWSA-N 0.000 description 1
- YKBSXQFZWFXFIB-VOAKCMCISA-N Lys-Thr-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCCN)C(O)=O YKBSXQFZWFXFIB-VOAKCMCISA-N 0.000 description 1
- RQILLQOQXLZTCK-KBPBESRZSA-N Lys-Tyr-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O RQILLQOQXLZTCK-KBPBESRZSA-N 0.000 description 1
- PPNCMJARTHYNEC-MEYUZBJRSA-N Lys-Tyr-Thr Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)CC1=CC=C(O)C=C1 PPNCMJARTHYNEC-MEYUZBJRSA-N 0.000 description 1
- DCHHUGLTVLJYKA-FXQIFTODSA-N Met-Asn-Ala Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O DCHHUGLTVLJYKA-FXQIFTODSA-N 0.000 description 1
- DBOMZJOESVYERT-GUBZILKMSA-N Met-Asn-Met Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCSC)C(=O)O)N DBOMZJOESVYERT-GUBZILKMSA-N 0.000 description 1
- OOSPRDCGTLQLBP-NHCYSSNCSA-N Met-Glu-Val Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OOSPRDCGTLQLBP-NHCYSSNCSA-N 0.000 description 1
- AWGBEIYZPAXXSX-RWMBFGLXSA-N Met-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCSC)N AWGBEIYZPAXXSX-RWMBFGLXSA-N 0.000 description 1
- XDGFFEZAZHRZFR-RHYQMDGZSA-N Met-Leu-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XDGFFEZAZHRZFR-RHYQMDGZSA-N 0.000 description 1
- ZRACLHJYVRBJFC-ULQDDVLXSA-N Met-Lys-Phe Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 ZRACLHJYVRBJFC-ULQDDVLXSA-N 0.000 description 1
- ATBJCCFCJXCNGZ-UFYCRDLUSA-N Met-Tyr-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 ATBJCCFCJXCNGZ-UFYCRDLUSA-N 0.000 description 1
- 241000204031 Mycoplasma Species 0.000 description 1
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 235000008098 Oxalis acetosella Nutrition 0.000 description 1
- 241001119522 Paullinia pinnata Species 0.000 description 1
- 235000010240 Paullinia pinnata Nutrition 0.000 description 1
- LDSOBEJVGGVWGD-DLOVCJGASA-N Phe-Asp-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 LDSOBEJVGGVWGD-DLOVCJGASA-N 0.000 description 1
- CSYVXYQDIVCQNU-QWRGUYRKSA-N Phe-Asp-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O CSYVXYQDIVCQNU-QWRGUYRKSA-N 0.000 description 1
- SXJGROGVINAYSH-AVGNSLFASA-N Phe-Gln-Asp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N SXJGROGVINAYSH-AVGNSLFASA-N 0.000 description 1
- MGBRZXXGQBAULP-DRZSPHRISA-N Phe-Glu-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 MGBRZXXGQBAULP-DRZSPHRISA-N 0.000 description 1
- KJJROSNFBRWPHS-JYJNAYRXSA-N Phe-Glu-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O KJJROSNFBRWPHS-JYJNAYRXSA-N 0.000 description 1
- MYQCCQSMKNCNKY-KKUMJFAQSA-N Phe-His-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)N[C@@H](CO)C(=O)O)N MYQCCQSMKNCNKY-KKUMJFAQSA-N 0.000 description 1
- JQLQUPIYYJXZLJ-ZEWNOJEFSA-N Phe-Ile-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 JQLQUPIYYJXZLJ-ZEWNOJEFSA-N 0.000 description 1
- KXUZHWXENMYOHC-QEJZJMRPSA-N Phe-Leu-Ala Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O KXUZHWXENMYOHC-QEJZJMRPSA-N 0.000 description 1
- RSPUIENXSJYZQO-JYJNAYRXSA-N Phe-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 RSPUIENXSJYZQO-JYJNAYRXSA-N 0.000 description 1
- WLYPRKLMRIYGPP-JYJNAYRXSA-N Phe-Lys-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 WLYPRKLMRIYGPP-JYJNAYRXSA-N 0.000 description 1
- YOFKMVUAZGPFCF-IHRRRGAJSA-N Phe-Met-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(O)=O YOFKMVUAZGPFCF-IHRRRGAJSA-N 0.000 description 1
- RYQWALWYQWBUKN-FHWLQOOXSA-N Phe-Phe-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O RYQWALWYQWBUKN-FHWLQOOXSA-N 0.000 description 1
- CKJACGQPCPMWIT-UFYCRDLUSA-N Phe-Pro-Phe Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CKJACGQPCPMWIT-UFYCRDLUSA-N 0.000 description 1
- MVIJMIZJPHQGEN-IHRRRGAJSA-N Phe-Ser-Val Chemical compound CC(C)[C@@H](C([O-])=O)NC(=O)[C@H](CO)NC(=O)[C@@H]([NH3+])CC1=CC=CC=C1 MVIJMIZJPHQGEN-IHRRRGAJSA-N 0.000 description 1
- XNQMZHLAYFWSGJ-HTUGSXCWSA-N Phe-Thr-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O XNQMZHLAYFWSGJ-HTUGSXCWSA-N 0.000 description 1
- MJOJSHOTYWABPR-WIRXVTQYSA-N Phe-Trp-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=CC=C1 MJOJSHOTYWABPR-WIRXVTQYSA-N 0.000 description 1
- BAONJAHBAUDJKA-BZSNNMDCSA-N Phe-Tyr-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=CC=C1 BAONJAHBAUDJKA-BZSNNMDCSA-N 0.000 description 1
- QUUCAHIYARMNBL-FHWLQOOXSA-N Phe-Tyr-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N QUUCAHIYARMNBL-FHWLQOOXSA-N 0.000 description 1
- FRMKIPSIZSFTTE-HJOGWXRNSA-N Phe-Tyr-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O FRMKIPSIZSFTTE-HJOGWXRNSA-N 0.000 description 1
- KUSYCSMTTHSZOA-DZKIICNBSA-N Phe-Val-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N KUSYCSMTTHSZOA-DZKIICNBSA-N 0.000 description 1
- APZNYJFGVAGFCF-JYJNAYRXSA-N Phe-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)Cc1ccccc1)C(C)C)C(O)=O APZNYJFGVAGFCF-JYJNAYRXSA-N 0.000 description 1
- VCYJKOLZYPYGJV-AVGNSLFASA-N Pro-Arg-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O VCYJKOLZYPYGJV-AVGNSLFASA-N 0.000 description 1
- WFHYFCWBLSKEMS-KKUMJFAQSA-N Pro-Glu-Phe Chemical compound N([C@@H](CCC(=O)O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C(=O)[C@@H]1CCCN1 WFHYFCWBLSKEMS-KKUMJFAQSA-N 0.000 description 1
- SXMSEHDMNIUTSP-DCAQKATOSA-N Pro-Lys-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O SXMSEHDMNIUTSP-DCAQKATOSA-N 0.000 description 1
- PUQRDHNIOONJJN-AVGNSLFASA-N Pro-Lys-Met Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(O)=O PUQRDHNIOONJJN-AVGNSLFASA-N 0.000 description 1
- ULWBBFKQBDNGOY-RWMBFGLXSA-N Pro-Lys-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCCCN)C(=O)N2CCC[C@@H]2C(=O)O ULWBBFKQBDNGOY-RWMBFGLXSA-N 0.000 description 1
- WCNVGGZRTNHOOS-ULQDDVLXSA-N Pro-Lys-Tyr Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O WCNVGGZRTNHOOS-ULQDDVLXSA-N 0.000 description 1
- QGLFRQCECIWXFA-RCWTZXSCSA-N Pro-Met-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@@H]1CCCN1)O QGLFRQCECIWXFA-RCWTZXSCSA-N 0.000 description 1
- ZUZINZIJHJFJRN-UBHSHLNASA-N Pro-Phe-Ala Chemical compound C([C@@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 ZUZINZIJHJFJRN-UBHSHLNASA-N 0.000 description 1
- VGVCNKSUVSZEIE-IHRRRGAJSA-N Pro-Phe-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O VGVCNKSUVSZEIE-IHRRRGAJSA-N 0.000 description 1
- JIWJRKNYLSHONY-KKUMJFAQSA-N Pro-Phe-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JIWJRKNYLSHONY-KKUMJFAQSA-N 0.000 description 1
- UIUWGMRJTWHIJZ-ULQDDVLXSA-N Pro-Tyr-Lys Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CCCCN)C(=O)O UIUWGMRJTWHIJZ-ULQDDVLXSA-N 0.000 description 1
- IYCBDVBJWDXQRR-FXQIFTODSA-N Ser-Ala-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O IYCBDVBJWDXQRR-FXQIFTODSA-N 0.000 description 1
- HBZBPFLJNDXRAY-FXQIFTODSA-N Ser-Ala-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O HBZBPFLJNDXRAY-FXQIFTODSA-N 0.000 description 1
- KNZQGAUEYZJUSQ-ZLUOBGJFSA-N Ser-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CO)N KNZQGAUEYZJUSQ-ZLUOBGJFSA-N 0.000 description 1
- OJPHFSOMBZKQKQ-GUBZILKMSA-N Ser-Gln-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CO OJPHFSOMBZKQKQ-GUBZILKMSA-N 0.000 description 1
- QMCDMHWAKMUGJE-IHRRRGAJSA-N Ser-Phe-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O QMCDMHWAKMUGJE-IHRRRGAJSA-N 0.000 description 1
- GZGFSPWOMUKKCV-NAKRPEOUSA-N Ser-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO GZGFSPWOMUKKCV-NAKRPEOUSA-N 0.000 description 1
- FLMYSKVSDVHLEW-SVSWQMSJSA-N Ser-Thr-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FLMYSKVSDVHLEW-SVSWQMSJSA-N 0.000 description 1
- RTXKJFWHEBTABY-IHPCNDPISA-N Ser-Trp-Tyr Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)O)NC(=O)[C@H](CO)N RTXKJFWHEBTABY-IHPCNDPISA-N 0.000 description 1
- GSCVDSBEYVGMJQ-SRVKXCTJSA-N Ser-Tyr-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CO)N)O GSCVDSBEYVGMJQ-SRVKXCTJSA-N 0.000 description 1
- 101710158453 Sex-specific storage-protein 2 Proteins 0.000 description 1
- TYVAWPFQYFPSBR-BFHQHQDPSA-N Thr-Ala-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)NCC(O)=O TYVAWPFQYFPSBR-BFHQHQDPSA-N 0.000 description 1
- TZKPNGDGUVREEB-FOHZUACHSA-N Thr-Asn-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O TZKPNGDGUVREEB-FOHZUACHSA-N 0.000 description 1
- JTEICXDKGWKRRV-HJGDQZAQSA-N Thr-Asn-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N)O JTEICXDKGWKRRV-HJGDQZAQSA-N 0.000 description 1
- ZUUDNCOCILSYAM-KKHAAJSZSA-N Thr-Asp-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O ZUUDNCOCILSYAM-KKHAAJSZSA-N 0.000 description 1
- WLDUCKSCDRIVLJ-NUMRIWBASA-N Thr-Gln-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O WLDUCKSCDRIVLJ-NUMRIWBASA-N 0.000 description 1
- LIXBDERDAGNVAV-XKBZYTNZSA-N Thr-Gln-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O LIXBDERDAGNVAV-XKBZYTNZSA-N 0.000 description 1
- XHWCDRUPDNSDAZ-XKBZYTNZSA-N Thr-Ser-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N)O XHWCDRUPDNSDAZ-XKBZYTNZSA-N 0.000 description 1
- AHERARIZBPOMNU-KATARQTJSA-N Thr-Ser-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O AHERARIZBPOMNU-KATARQTJSA-N 0.000 description 1
- CSNBWOJOEOPYIJ-UVOCVTCTSA-N Thr-Thr-Lys Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(O)=O CSNBWOJOEOPYIJ-UVOCVTCTSA-N 0.000 description 1
- QUILOGWWLXMSAT-IHRRRGAJSA-N Tyr-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QUILOGWWLXMSAT-IHRRRGAJSA-N 0.000 description 1
- XQYHLZNPOTXRMQ-KKUMJFAQSA-N Tyr-Glu-Arg Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O XQYHLZNPOTXRMQ-KKUMJFAQSA-N 0.000 description 1
- GIOBXJSONRQHKQ-RYUDHWBXSA-N Tyr-Gly-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O GIOBXJSONRQHKQ-RYUDHWBXSA-N 0.000 description 1
- FNWGDMZVYBVAGJ-XEGUGMAKSA-N Tyr-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC1=CC=C(C=C1)O)N FNWGDMZVYBVAGJ-XEGUGMAKSA-N 0.000 description 1
- USYGMBIIUDLYHJ-GVARAGBVSA-N Tyr-Ile-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 USYGMBIIUDLYHJ-GVARAGBVSA-N 0.000 description 1
- GYKDRHDMGQUZPU-MGHWNKPDSA-N Tyr-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC1=CC=C(C=C1)O)N GYKDRHDMGQUZPU-MGHWNKPDSA-N 0.000 description 1
- SINRIKQYQJRGDQ-MEYUZBJRSA-N Tyr-Lys-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 SINRIKQYQJRGDQ-MEYUZBJRSA-N 0.000 description 1
- NVZVJIUDICCMHZ-BZSNNMDCSA-N Tyr-Phe-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O NVZVJIUDICCMHZ-BZSNNMDCSA-N 0.000 description 1
- SOAUMCDLIUGXJJ-SRVKXCTJSA-N Tyr-Ser-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O SOAUMCDLIUGXJJ-SRVKXCTJSA-N 0.000 description 1
- MWUYSCVVPVITMW-IGNZVWTISA-N Tyr-Tyr-Ala Chemical compound C([C@@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 MWUYSCVVPVITMW-IGNZVWTISA-N 0.000 description 1
- TYGHOWWWMTWVKM-HJOGWXRNSA-N Tyr-Tyr-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 TYGHOWWWMTWVKM-HJOGWXRNSA-N 0.000 description 1
- KHPLUFDSWGDRHD-SLFFLAALSA-N Tyr-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N)C(=O)O KHPLUFDSWGDRHD-SLFFLAALSA-N 0.000 description 1
- RMRFSFXLFWWAJZ-HJOGWXRNSA-N Tyr-Tyr-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 RMRFSFXLFWWAJZ-HJOGWXRNSA-N 0.000 description 1
- ABSXSJZNRAQDDI-KJEVXHAQSA-N Tyr-Val-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ABSXSJZNRAQDDI-KJEVXHAQSA-N 0.000 description 1
- JIODCDXKCJRMEH-NHCYSSNCSA-N Val-Arg-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N JIODCDXKCJRMEH-NHCYSSNCSA-N 0.000 description 1
- IVXJODPZRWHCCR-JYJNAYRXSA-N Val-Arg-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N IVXJODPZRWHCCR-JYJNAYRXSA-N 0.000 description 1
- BMGOFDMKDVVGJG-NHCYSSNCSA-N Val-Asp-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BMGOFDMKDVVGJG-NHCYSSNCSA-N 0.000 description 1
- YODDULVCGFQRFZ-ZKWXMUAHSA-N Val-Asp-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O YODDULVCGFQRFZ-ZKWXMUAHSA-N 0.000 description 1
- FOADDSDHGRFUOC-DZKIICNBSA-N Val-Glu-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N FOADDSDHGRFUOC-DZKIICNBSA-N 0.000 description 1
- WNZSAUMKZQXHNC-UKJIMTQDSA-N Val-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N WNZSAUMKZQXHNC-UKJIMTQDSA-N 0.000 description 1
- UMPVMAYCLYMYGA-ONGXEEELSA-N Val-Leu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O UMPVMAYCLYMYGA-ONGXEEELSA-N 0.000 description 1
- QRVPEKJBBRYISE-XUXIUFHCSA-N Val-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C(C)C)N QRVPEKJBBRYISE-XUXIUFHCSA-N 0.000 description 1
- MJOUSKQHAIARKI-JYJNAYRXSA-N Val-Phe-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)CC1=CC=CC=C1 MJOUSKQHAIARKI-JYJNAYRXSA-N 0.000 description 1
- LGXUZJIQCGXKGZ-QXEWZRGKSA-N Val-Pro-Asn Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)N)C(=O)O)N LGXUZJIQCGXKGZ-QXEWZRGKSA-N 0.000 description 1
- QIVPZSWBBHRNBA-JYJNAYRXSA-N Val-Pro-Phe Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1ccccc1)C(O)=O QIVPZSWBBHRNBA-JYJNAYRXSA-N 0.000 description 1
- ZLNYBMWGPOKSLW-LSJOCFKGSA-N Val-Val-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLNYBMWGPOKSLW-LSJOCFKGSA-N 0.000 description 1
- AOILQMZPNLUXCM-AVGNSLFASA-N Val-Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN AOILQMZPNLUXCM-AVGNSLFASA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 108010045023 alanyl-prolyl-tyrosine Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 108010070783 alanyltyrosine Proteins 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010073628 glutamyl-valyl-phenylalanine Proteins 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 108010092114 histidylphenylalanine Proteins 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 108010051673 leucyl-glycyl-phenylalanine Proteins 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 1
- 108010030617 leucyl-phenylalanyl-valine Proteins 0.000 description 1
- 108010091871 leucylmethionine Proteins 0.000 description 1
- 108010044348 lysyl-glutamyl-aspartic acid Proteins 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- -1 metals ion Chemical class 0.000 description 1
- 108010068488 methionylphenylalanine Proteins 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 108010073101 phenylalanylleucine Proteins 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 108010077037 tyrosyl-tyrosyl-phenylalanine Proteins 0.000 description 1
- 108010003137 tyrosyltyrosine Proteins 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 108010009962 valyltyrosine Proteins 0.000 description 1
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a preparation method for natural sex storage protein-2 and a use for the same, belonging to the field of biotechnology. The preparation method disclosed by the invention comprises the following steps of: using silkworm chrysalis as a raw material, homogenizing, filtering, centrifuging and collecting the supernatant, and then separating and purifying by the intrinsic characteristic of high temperature resistance of protein to obtain a high-purity natural sex storage protein-2 (SSP2), and using the same for cell culture. The method provided by the invention is easy to get raw material, low in production cost, simple and easy in operation, high in yield and suitable for large-scale production; additionally, the purified sex storage protein-2 can be used as a protective agent in cell culture for protecting cells from apoptosis due to cell stress.
Description
Technical field
The present invention relates to a kind of natural sex and store white 2, belong to biological technical field.
Background technology
Silkworm chrysalis is as unique insects food in " as the new resource for food list of bread and cheese management " of Ministry of Health's approval.It has high nutritive value, contains rich in protein, lipid acid and VITAMINs.(sex-specific storage-protein 2 SSP2), shows through cell experiment wherein contained sex storage protein 2, has the protection cell, anti-apoptotic, somatotrophic effect.At present, Ox blood serum is necessary in the cell cultivation process, contains the abundant necessary nutritive ingredient of cell growth; Provide conjugated protein; Discern VITAMINs, lipid, metals ion, and combine, play detoxification with toxic metal box heat source substance; The effect of potential of hydrogen damping fluid is played in the source that is cell attachment, sprawls the required factor of growth.But also there is more hidden danger in Ox blood serum, so its specification of quality is very strict, must prove cows or the country from no mad cow disease, does not contain the inhibition to production vaccine virus, and does not have ground contaminations such as mycoplasma, virus.And sex storage protein 2 preparing methods are simple, need not to consider the interference of above-mentioned pathogenic bacteria etc., and being expected to develop becomes the product that substitutes Ox blood serum, therefore has great Application Research prospect.
Summary of the invention
In view of this, the purpose of this invention is to provide a kind of natural sex and store white 2.The present invention as raw material, utilizes the resistant to elevated temperatures characteristic of this albumen with silkworm chrysalis, obtains to have high biological activity through simple separation and purification means; Highly purified sex storage protein 2; And be used for cell cultures with its substitute blood serum, and it is easy and simple to handle, and technical requirements is not high; Output is big, is applicable to scale operation.
For achieving the above object, the present invention provides a kind of natural sex storage protein 2, and its aminoacid sequence is shown in SEQ ID NO:1.
The preparation method of above-mentioned natural sex storage protein 2 may further comprise the steps:
(1) silkworm chrysalis sample pretreatment: with the 20-50mM phosphoric acid buffer or the zero(ppm) water of fresh silkworm chrysalis and 4-20 ℃ of precooling; Joined in the juicer homogenate 30 seconds by the mass volume ratio of 1:1,, continue homogenate ice bath on ice 5 minutes; Repeat 3 times, the albumen in the silkworm chrysalis is fully dissolved; In Centrifuge Cup, 4 ℃, 8000-15000 rev/min, centrifugal 20-30 minute, supernatant passes through nine layers of filtered through gauze except that behind the degrease, and was centrifugal, repeated centrifugation and filtration step 3-5 time with the homogenate sample transfer; Collect supernatant, 4 ℃, 8000-15000 rev/min, centrifugal 20-30 minute, supernatant was with 0.22 μ m or 0.45 μ m filter membrane vacuum filtration, 4 ℃ of preservations;
(2) initial gross separation purifying: the supernatant that step (1) pre-treatment is obtained places 50 ℃ of waters bath with thermostatic control 30 minutes, and 8000-15000 rev/min again, centrifugal 20-30 minute; Get supernatant, nine layers of filtered through gauze repeat 50 ℃ of water-baths; Centrifugal and filtration step 3 times are collected supernatant to place≤80 ℃ of waters bath with thermostatic control 30 minutes 8000-15000 rev/min again; Centrifugal 20-30 minute, get supernatant, nine layers of filtered through gauze; Same step repeats (the same temperature of experiment proof repeats 3 abilities and fully removes other foreign proteins, but liquor capacity need increase number of times greater than 2L) 3-5 time;
(3) further separation and purification: the supernatant that step (2) initial gross separation purifying is obtained is with the ultrafiltration of 100kD ultra-filtration membrane bag; Filtrate continues with the ultrafiltration of 10kD ultra-filtration membrane bag; To get greater than the filtrate sample of 100kD and use Superdex greater than 100kD, greater than 10kD, less than 100kD and subsequent use respectively less than 4 ℃ of preservations of filtrate of 10kD
TM200 separate, and the collection back, peak that will contain target protein concentrates, and with the further separation and purification of Resource Q post, can obtain said natural sex storage protein 2.
Preferably, the acidity of phosphoric acid buffer is pH7.4-8.0 in the above-mentioned steps (1).
The present invention also provides the purposes of a kind of above-mentioned natural sex storage protein 2 in cell cultures, and after the natural sex storage protein 2 usefulness 30kD ultrafiltration pipes that above-mentioned further separation and purification is obtained concentrated quantitatively, 4 ℃ subsequent use; Prepare the cell culture medium that does not contain serum but contain the natural sex storage protein 2 of 25-1000 μ g/mL then, concrete steps are following: sex storage protein 2 is carried out concentration determination; Cross 0.2 μ m or 0.22 μ m filter degerming after being diluted to certain concentration with serum free medium again; Cell needs to wash gently 2 times with serum free medium; The substratum that contains sex storage protein 2 for preparing is joined in the cell, and pair cell is cultivated; Cultivate after five days, adopt mtt assay to detect cytoactive.
Preferably, the acidity of phosphoric acid buffer is pH7.4-8.0 in the above-mentioned steps (1).
The present invention also provides the purposes of above-mentioned natural sex storage protein 2 in cell cultures.
Preferably, after above-mentioned natural sex storage protein 2 usefulness 30kD ultrafiltration pipes concentrated quantitatively, 4 ℃ subsequent use; Prepare the cell culture medium that does not contain serum but contain the natural sex storage protein 2 of 25-100 μ g/mL then, BmN cell, Hek293 cell are cultivated, and detected cytoactive with mtt assay.
The present invention can utilize silkworm chrysalis as raw material; Obtain to have resistant to elevated temperatures sex storage protein through separation and purification, this method is easy and simple to handle, and output is high; Be applicable to scale operation; The sex storage protein of purifying can be used as the protective material in the cell cultures, and the protection cell is in order to avoid cellular stress and cause apoptosis, and has no side effect.
Description of drawings
Fig. 1 is the electrophoretic analysis figure of natural sex storage protein 2 separation and purification; M: low molecular mass protein standard; 1: supernatant total protein after the dried silkworm chrysalis meal homogenate; 2:50 ℃ of water-bath 30min supernatant; 3:50 ℃ of water-bath 30min deposition; 4:60 ℃ of water-bath 30min supernatant; 5:60 ℃ of water-bath 30min deposition; 6:70 ℃ of water-bath 30min supernatant; 7:70 ℃ of water-bath 30min deposition; 8:80 ℃ of water-bath 30min supernatant; 9:80 ℃ of water-bath 30min deposition;
Fig. 2 is the Superdex of natural sex storage protein 2
TM200 analyze color atlas;
Fig. 3 is Superdex among Fig. 2
TMThe electrophoretic analysis figure of 200 separating natural sex storage proteins 2; M: low molecular mass protein standard; 1: former state (reductibility sample-loading buffer); 2: 1# peak among Fig. 2 (reductibility sample-loading buffer); 3: former state (irreducibility sample-loading buffer); 4: 1# peak among Fig. 2 (irreducibility sample-loading buffer);
Fig. 4 is the further analysis color atlas of Resource Q post; Wherein, 1# peak: not conjugated protein; 2# peak: 280mM NaCl eluted protein;
Fig. 5 is the electrophoretic analysis figure at 2# peak among Fig. 4;
Fig. 6 is the evaluation mass spectrum of natural sex storage protein 2;
Fig. 7 cultivates cell death inducing test MTT interpretation of result figure for the BmN cell non-serum; Wherein, no F is a serum-free, and 25 μ g/mL, 50 μ g/mL, 100 μ g/mL, 200 μ g/mL, 500 μ g/mL, 1000 μ g/mL are respectively the final concentration of natural sex storage protein 2;
Fig. 8 cultivates cell death inducing test MTT interpretation of result figure for the HEK293 cell non-serum; Wherein, no F is a serum-free, and 25 μ g/mL, 50 μ g/mL, 100 μ g/mL, 200 μ g/mL, 500 μ g/mL, 1000 μ g/mL are respectively the final concentration of natural sex storage protein 2.
Embodiment
Be noted that following specifying all is exemplary, being intended to provides further invention to the present invention.Except as otherwise noted, all Science and Technology terms of using of this paper have with the present invention under the identical meanings of person skilled common sense.
Set forth particular content of the present invention in detail below in conjunction with embodiment.
Embodiment 1:
The preparation method of a kind of natural sex storage protein 2 (SSP2) may further comprise the steps:
(1) silkworm chrysalis sample pretreatment: with the 50mM phosphoric acid buffer (pH7.4) of fresh silkworm chrysalis (available from Zhongqi Biological Pharmaceutical Co., Ltd., Zhejiang) with 4 ℃ of precoolings; Joined in the juicer homogenate 30 seconds by the mass volume ratio of 1:1; Ice bath on ice 5 minutes; Continue homogenate, repeat 3 times, the albumen in the silkworm chrysalis is fully dissolved; In Centrifuge Cup, 4 ℃, 15000 rev/mins, centrifugal 20 minutes, supernatant passes through nine layers of filtered through gauze except that behind the degrease, and was centrifugal, repeated centrifugation and filtration step 3 times with the homogenate sample transfer; Collect supernatant, 4 ℃, 15000 rev/mins, centrifugal 20 minutes, supernatant was with 0.45 μ m filter membrane vacuum filtration, 4 ℃ of preservations, and as shown in Figure 1, the protein electrophoresis figure that obtains is No. 1 swimming lane;
(2) initial gross separation purifying: the supernatant that pre-treatment is obtained places 50 ℃ of waters bath with thermostatic control 30 minutes, and 12000 rev/mins again, centrifugal 20 minutes, get supernatant, nine layers of filtered through gauze repeat 50 ℃ of water-baths, centrifugal and filtration step 3 times; Collect supernatant and place 80 ℃ of waters bath with thermostatic control 30 minutes, 15000 rev/mins again, centrifugal 20 minutes; Get supernatant, nine layers of filtered through gauze, same step repeats 3 times; It is subsequent use to collect 80 ℃ of 4 ℃ of pre-treatment supernatants; As shown in Figure 1, the protein electrophoresis figure that obtains is No. 8 swimming lane (explain that sex storage protein 2 is present in 80 ℃ of supernatants, the anti-80 ℃ of high temperature of sex storage protein also are described simultaneously);
(3) further separation and purification: the supernatant that step (2) initial gross separation purifying is obtained is with the ultrafiltration of 100kD ultra-filtration membrane bag; Filtrate continues with the ultrafiltration of 10kD ultra-filtration membrane bag, respectively will be greater than 100kD, greater than 10kD, less than 100kD and subsequent use less than 4 ℃ of preservations of filtrate of 10kD; Get greater than the filtrate sample of 100kD and use Superdex
TM200 separate, and the collection back, peak that will contain target protein concentrates 1# marker peak promptly shown in Figure 2; Its electrophorogram is (band of arrow indication position is the electrophoretic band of sex storage protein 2) shown in accompanying drawing 3, and with the further separation and purification (see figure 4) of Resource Q post, and 2# peak among Fig. 4 is collected and carried out electrophoresis; Its electrophorogram (arrow indication position is the electrophoretic band of sex storage protein 2) as shown in Figure 5; Can obtain purer natural sex storage protein 2, the protein sample with this purifying obtains carries out mass spectrometric detection; Result such as accompanying drawing 6; The albumen fraction of coverage that is numbered Bmb013502 in this proteic peptide finger printing and the southwestern agricultural university silkworm albumen database reaches 53.9118%, and peptide section coupling number is 41 sections, and black character is concrete coupling peptide section in the square frame of Fig. 6 upper right corner; And obtain its aminoacid sequence shown in SEQ ID NO:1, prove that this albumen is natural sex storage protein 2.
The purposes of above-mentioned natural sex storage protein 2 in cell cultures: with above-mentioned steps (3) further natural sex storage protein 2 usefulness the 30kD ultrafiltration pipes that obtain of separation and purification concentrate quantitative after, 4 ℃ are subsequent use; Because of the homology of sex storage protein 2 and storage protein 2 reaches 90.65%; So design is to BmN cell (life science institute of Institutes Of Technology Of Zhejiang biochemical research is presented); The cytosis test of Hek293 cell (available from Beijing gold amethyst biological medicine technology ltd); Preparation does not contain serum; Contain the cell culture medium of the natural sex storage protein 2 of finite concentration (i.e. 25 μ g/mL-1000 μ g/mL), concrete steps are following: this natural sex storage protein 2 is carried out concentration determination; Using serum free medium (if BmN cell non-serum culture medium SF900 II is available from U.S. Gibico company, Hek293 cell non-serum culture medium DMEM available from Bioroc hundred gram medical biotechnology Ltd) to be diluted to final concentration respectively again is to cross 0.2 μ m filter degerming behind 25 μ g/mL, 50 μ g/mL, 100 μ g/mL, 200 μ g/mL, 500 μ g/mL, the 1000 μ g/mL; BmN cell and Hek293 cell need use BmN cell non-serum culture medium and Hek293 cell non-serum culture medium to wash gently 2 times; These substratum that contain this natural sex storage protein 2 that prepare are joined respectively in above-mentioned BmN cell and the Hek293 cell, these cells are cultivated; Cultivate after five days, adopt mtt assay to detect these cell activity.Experimental result proves; BmN cell and Hek293 cell activity are than high (like Fig. 7, shown in 8) in the serum free medium under the condition that natural sex storage protein 2 final concentrations are 50 μ g/mL; Can be applicable to cell cultures; Therefore the natural sex storage protein 2 that obtains of this separation and purification can be protected cell, has the effect of anti-apoptotic.
Embodiment 2:
The preparation method of a kind of natural sex storage protein 2 (SSP2) culturing cell may further comprise the steps:
(1) silkworm chrysalis sample pretreatment: with the zero(ppm) water of fresh silkworm chrysalis (available from Zhongqi Biological Pharmaceutical Co., Ltd., Zhejiang) with 20 ℃ of precoolings; Joined in the juicer homogenate 30 seconds by the mass volume ratio of 1:1; Ice bath on ice 5 minutes; Continue homogenate, repeat 3 times, the albumen in the silkworm chrysalis is fully dissolved; In Centrifuge Cup, 4 ℃, 8000 rev/mins, centrifugal 30 minutes, supernatant passes through nine layers of filtered through gauze except that behind the degrease, and was centrifugal, repeated centrifugation and filtration step 3 times with the homogenate sample transfer; Collect supernatant, 4 ℃, 8000 rev/mins, centrifugal 30 minutes, supernatant was with 0.45 μ m filter membrane vacuum filtration, 4 ℃ of preservations, and as shown in Figure 1, the protein electrophoresis figure that obtains is No. 1 swimming lane;
(2) initial gross separation purifying: the supernatant that pre-treatment is obtained places 50 ℃ of waters bath with thermostatic control 30 minutes, and 8000 rev/mins again, centrifugal 30 minutes, get supernatant, nine layers of filtered through gauze repeat 50 ℃ of water-baths, centrifugal and filtration step 3 times; Collect supernatant and place 80 ℃ of waters bath with thermostatic control 30 minutes, 8000 rev/mins again, centrifugal 30 minutes; Get supernatant, nine layers of filtered through gauze, same step repeats 3 times; It is subsequent use, as shown in Figure 1 to collect 80 ℃ of 4 ℃ of pre-treatment supernatants, and the protein electrophoresis figure that obtains is No. 8 swimming lane;
(3) further separation and purification: the supernatant that step (2) initial gross separation purifying is obtained is with the ultrafiltration of 100kD ultra-filtration membrane bag; Filtrate continues with the ultrafiltration of 10kD ultra-filtration membrane bag, respectively will be greater than 100kD, greater than 10kD, less than 100kD and subsequent use less than 4 ℃ of preservations of filtrate of 10kD; Get greater than the filtrate sample of 100kD and use Superdex
TM200 separate, and the collection back, peak that will contain target protein concentrates 1# marker peak promptly shown in Figure 2; Its electrophorogram and with the further separation and purification (see figure 4) of Resource Q post, is collected 2# peak among Fig. 4 and carry out electrophoresis shown in accompanying drawing 3; Its electrophorogram is as shown in Figure 5, can obtain purer natural sex storage protein 2, the protein sample that this purifying is obtained; Carry out mass spectrometric detection, the result is shown in accompanying drawing 6, and the albumen fraction of coverage that is numbered Bmb013502 in this proteic peptide finger printing and the southwestern agricultural university silkworm albumen database reaches 53.9118%; Peptide section coupling number is 41 sections; Black character is concrete coupling peptide section in the square frame of Fig. 6 upper right corner, and obtains its aminoacid sequence shown in SEQ ID NO:1, proves that this albumen is natural sex storage protein 2.
The purposes of above-mentioned natural sex storage protein 2 in cell cultures: with above-mentioned steps (3) further natural sex storage protein 2 usefulness the 30kD ultrafiltration pipes that obtain of separation and purification concentrate quantitative after, 4 ℃ are subsequent use; Because of the homology of sex storage protein 2 and storage protein 2 reaches 90.65%; So design is to BmN cell (life science institute of Institutes Of Technology Of Zhejiang biochemical research is presented); The cytosis test of Hek293 cell (available from Beijing gold amethyst biological medicine technology ltd); Preparation does not contain serum; Contain the cell culture medium of the natural sex storage protein 2 of finite concentration (i.e. 25 μ g/mL-1000 μ g/mL), concrete steps are following: this natural sex storage protein 2 is carried out concentration determination; Using serum free medium (if BmN cell non-serum culture medium SF900 II is available from U.S. Gibico company, Hek293 cell non-serum culture medium DMEM available from Bioroc hundred gram medical biotechnology Ltd) to be diluted to final concentration respectively again is to cross 0.22 μ m filter degerming behind 25 μ g/mL, 50 μ g/mL, 100 μ g/mL, 200 μ g/mL, 500 μ g/mL, the 1000 μ g/mL; BmN cell and Hek293 cell need use BmN cell non-serum culture medium and Hek293 cell non-serum culture medium to wash gently 2 times; These substratum that contain this natural sex storage protein 2 that prepare are joined respectively in above-mentioned BmN cell and the Hek293 cell, these cells are cultivated; Cultivate after five days, adopt mtt assay to detect these cell activity.Experimental result proves; BmN cell and Hek293 cell activity are than high in the serum free medium under the condition that natural sex storage protein 2 final concentrations are 50 μ g/mL; Can be applicable to cell cultures; Therefore the natural sex storage protein 2 that obtains of this separation and purification can be protected cell, has the effect of anti-apoptotic.
In a word; The present invention can utilize silkworm chrysalis as raw material, obtains to have resistant to elevated temperatures natural sex storage protein 2 through separation and purification, and the natural sex storage protein 2 of purifying can be used as the protective material in the cell cultures; The protection cell is in order to avoid cellular stress and cause apoptosis, and has no side effect; This method is easy and simple to handle, and the result is stable, and repetition rate height and output are high, are applicable to scale operation.
The above is merely the preferred embodiments of the present invention, it is pointed out that for the those of ordinary skill in the present technique, under the prerequisite that does not break away from core technology characteristic of the present invention, can also save and use Superdex in the step 3
TM200, the treatment process of Resource Q, the sample that separation and purification obtains does not influence the application in cell cultures.
SEQUENCE LISTING
< 110>Tianjin Yaoyu Biotechnology Co., Ltd.
< 120>natural sex storage protein 2
<130> 111707-I-CP-TJYU
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 703
<212> PRT
< 213>the sex storage protein 2
<400> 1
Met Lys Ser Val Leu Ile Leu Ala Gly Leu Val Ala Val Ala Leu Ser
1 5 10 15
Ser Ala Val Pro Lys Pro Ser Thr Ile Lys Thr Lys Asn Val Asp Ala
20 25 30
Val Phe Val Glu Lys Gln Lys Lys Ile Leu Ser Phe Phe Gln Asp Val
35 40 45
Ser Gln Leu Asn Thr Asp Asp Glu Tyr Tyr Lys Ile Gly Lys Asp Tyr
50 55 60
Asp Ile Glu Met Asn Met Asp Asn Tyr Thr Asn Lys Lys Ala Val Glu
65 70 75 80
Glu Phe Leu Lys Met Tyr Arg Thr Gly Phe Met Pro Lys Asn Leu Glu
85 90 95
Phe Ser Val Phe Tyr Asp Lys Met Arg Asp Glu Ala Ile Ala Leu Phe
100 105 110
His Leu Phe Tyr Tyr Ala Lys Asp Phe Glu Thr Phe Tyr Lys Thr Ala
115 120 125
Cys Phe Ala Arg Val His Leu Asn Gln Gly Gln Phe Leu Tyr Ala Phe
130 135 140
Tyr Ile Ala Val Ile Gln Arg Ser Asp Cys His Gly Phe Val Val Pro
145 150 155 160
Ala Pro Tyr Glu Val Tyr Pro Lys Met Phe Met Asn Met Glu Val Leu
165 170 175
Gln Lys Ile Tyr Val Thr Lys Met Gln Asp Gly Leu Ile Asn Pro Glu
180 185 190
Ala Ala Ala Lys Tyr Gly Ile His Lys Glu Asn Asp Tyr Phe Val Tyr
195 200 205
Lys Ala Asn Tyr Ser Asn Ala Val Leu Tyr Asn Asn Glu Glu Gln Arg
210 215 220
Leu Thr Tyr Phe Thr Glu Asp Ile Gly Met Asn Ala Tyr Tyr Tyr Tyr
225 230 235 240
Phe His Ser His Leu Pro Phe Trp Trp Thr Ser Glu Lys Tyr Gly Ala
245 250 255
Leu Lys Glu Arg Arg Gly Glu Val Tyr Phe Tyr Phe Tyr Gln Gln Leu
260 265 270
Leu Ala Arg Tyr Tyr Phe Glu Arg Leu Thr Asn Gly Leu Gly Lys Ile
275 280 285
Pro Glu Phe Ser Trp Tyr Ser Pro Ile Lys Thr Gly Tyr Tyr Pro Leu
290 295 300
Met Leu Thr Lys Phe Thr Pro Phe Ala Gln Arg Pro Asp Tyr Tyr Asn
305 310 315 320
Leu His Thr Glu Glu Asn Tyr Glu Arg Val Arg Phe Leu Asp Thr Tyr
325 330 335
Glu Lys Thr Phe Val Gln Phe Leu Gln Lys Asp His Phe Glu Ala Phe
340 345 350
Gly Gln Lys Ile Asp Phe His Asp Pro Lys Ala Ile Asn Phe Val Gly
355 360 365
Asn Tyr Trp Gln Asp Asn Ala Asp Leu Tyr Gly Glu Glu Val Thr Lys
370 375 380
Asp Tyr Gln Arg Ser Tyr Glu Val Phe Ala Arg Arg Val Leu Gly Ala
385 390 395 400
Ala Pro Met Pro Phe Asp Lys Tyr Thr Phe Met Pro Ser Ala Met Asp
405 410 415
Phe Tyr Gln Thr Ser Leu Arg Asp Pro Ala Phe Tyr Gln Leu Tyr Asn
420 425 430
Arg Ile Val Glu Tyr Ile Val Glu Phe Lys Gln Tyr Leu Lys Pro Tyr
435 440 445
Thr Gln Asp Lys Leu Tyr Phe Asp Gly Val Lys Ile Thr Asp Val Lys
450 455 460
Val Asp Lys Leu Thr Thr Phe Phe Glu Asn Phe Glu Phe Asp Ala Ser
465 470 475 480
Asn Ser Val Tyr Phe Ser Lys Glu Glu Ile Lys Asn Asn His Val His
485 490 495
Asp Val Lys Val Arg Gln Pro Arg Leu Asn His Ser Pro Phe Asn Val
500 505 510
Asn Ile Glu Val Asp Ser Asn Val Ala Ser Asp Ala Val Val Lys Ile
515 520 525
Phe Leu Ala Pro Lys Tyr Asp Asp Asn Gly Ile Pro Leu Thr Leu Glu
530 535 540
Asp Asn Trp Met Lys Phe Phe Glu Leu Asp Trp Phe Thr Thr Lys Leu
545 550 555 560
Thr Ala Gly Gln Asn Lys Ile Ile Arg Asn Ser Asn Glu Phe Val Ile
565 570 575
Phe Lys Glu Asp Ser Val Pro Met Thr Glu Ile Met Lys Met Leu Asp
580 585 590
Glu Gly Lys Val Pro Phe Asp Met Ser Glu Glu Phe Cys Tyr Met Pro
595 600 605
Lys Arg Leu Met Leu Pro Arg Gly Thr Glu Gly Gly Phe Pro Phe Gln
610 615 620
Leu Phe Val Phe Val Tyr Pro Phe Asp Asn Lys Gly Lys Asp Leu Ala
625 630 635 640
Pro Phe Glu Ser Phe Val Leu Asp Asn Lys Pro Leu Gly Phe Pro Leu
645 650 655
Asp Arg Pro Val Val Asp Ala Leu Phe Lys Val Pro Asn Met Tyr Phe
660 665 670
Lys Asp Ile Phe Ile Tyr His Glu Gly Glu Arg Phe Pro Tyr Lys Phe
675 680 685
Asn Ile Pro Ser Tyr Asp Thr Gln Ser Asn Val Val Pro Lys Asn
690 695 700
Claims (5)
1. natural sex storage protein 2, its aminoacid sequence is shown in SEQ ID NO:1.
2. the preparation method of the said natural sex storage protein 2 of claim 1 is characterized in that, said method comprising the steps of:
(1) silkworm chrysalis sample pretreatment: with the 20-50mM phosphoric acid buffer or the zero(ppm) water of fresh silkworm chrysalis and 4 ℃ of-20 ℃ of precoolings; Joined in the juicer homogenate 30 seconds by the mass volume ratio of 1:1,, continue homogenate ice bath on ice 5 minutes; Repeat 3 times, the albumen in the silkworm chrysalis is fully dissolved; In Centrifuge Cup, 4 ℃, 8000-15000 rev/min, centrifugal 20-30 minute, supernatant passes through nine layers of filtered through gauze except that behind the degrease, and was centrifugal, repeated centrifugation and filtration step 3 times with the homogenate sample transfer; Collect supernatant, 4 ℃, 8000-15000 rev/min, centrifugal 20-30 minute, supernatant was with 0.22 μ m or 0.45 μ m filter membrane vacuum filtration, 4 ℃ of preservations;
(2) initial gross separation purifying: the supernatant that step (1) pre-treatment is obtained places 50 ℃ of waters bath with thermostatic control 30 minutes, 8000-15000 rev/min, centrifugal 20-30 minute, gets supernatant; Nine layers of filtered through gauze repeat 50 ℃ of water-baths, and centrifugal and filtration step 3-5 time, collection supernatant place≤80 ℃ of waters bath with thermostatic control 30 minutes; 8000-15000 rev/min again, centrifugal 20-30 minute, get supernatant; Nine layers of filtered through gauze, same step repeats 3-5 time, does not have deposition until 80 ℃ of waters bath with thermostatic control and separates out;
(3) further separation and purification: the supernatant that step (2) initial gross separation purifying is obtained will be subsequent use greater than 4 ℃ of preservations of liquid concentrator of 100kD with 100kD ultra-filtration membrane bag ultrafiltration and concentration, get greater than the liquid concentrator liquid sample of 100kD and use Superdex
TM200 separate, and the collection back, peak that will contain target protein concentrates, and with the further separation and purification of Resource Q post, can obtain said natural sex storage protein 2.
3. method according to claim 1 is characterized in that, the acidity of phosphoric acid buffer is p7.4-8.0 in the said step (1).
4. the purposes of the said natural sex storage protein 2 of claim 1 in cell cultures.
5. purposes according to claim 3 is characterized in that, concentrate the described natural sex storage protein 2 usefulness 30kD ultrafiltration pipes of claim 1 quantitatively after, 4 ℃ are subsequent use; Prepare the cell culture medium that does not contain serum but contain the natural sex storage protein 2 of 25-100 μ g/mL then, BmN cell, Hek293 cell are cultivated, and detected cytoactive with mtt assay.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110426497XA CN102516381B (en) | 2011-12-19 | 2011-12-19 | Natural sex storage protein-2 as well as preparation and use for same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110426497XA CN102516381B (en) | 2011-12-19 | 2011-12-19 | Natural sex storage protein-2 as well as preparation and use for same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102516381A true CN102516381A (en) | 2012-06-27 |
| CN102516381B CN102516381B (en) | 2013-10-30 |
Family
ID=46287494
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110426497XA Expired - Fee Related CN102516381B (en) | 2011-12-19 | 2011-12-19 | Natural sex storage protein-2 as well as preparation and use for same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102516381B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014075443A1 (en) * | 2012-11-13 | 2014-05-22 | 天津耀宇生物技术有限公司 | Purification method of protein group of 30 kd in silkworm pupa |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974545A (en) * | 2010-09-14 | 2011-02-16 | 浙江大学 | Method for massively expressing wild silkworm anti-viral protein SP-2 |
-
2011
- 2011-12-19 CN CN201110426497XA patent/CN102516381B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974545A (en) * | 2010-09-14 | 2011-02-16 | 浙江大学 | Method for massively expressing wild silkworm anti-viral protein SP-2 |
Non-Patent Citations (4)
| Title |
|---|
| EUN JEONG KIM ET AL.: "Isolation and Characterization of an Apotosis-inhibiting Component from the Hemolymph of Bombyx mori", 《BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS》 * |
| WON JONG RHEE ET AL.: "Inhibition of Hela Cell Apotosis by Storage-Protein 2", 《BIOTECHNOL. PROG.》 * |
| YAO H ET AL.: "Accession number: NP_001037590", 《GENBANK》 * |
| 马聪等: "猪脂肪GLUT4蛋白的大量提取纯化及鉴定", 《现代生物医学进展》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014075443A1 (en) * | 2012-11-13 | 2014-05-22 | 天津耀宇生物技术有限公司 | Purification method of protein group of 30 kd in silkworm pupa |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102516381B (en) | 2013-10-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102146135A (en) | Recombinant human-like collagen and production method thereof | |
| CN106810602A (en) | Protein and its application that arch insect infection Mice brain tissues differential expression protein and brain development regulatory protein interact | |
| CN102731641B (en) | Modification of echinococcus granulosus EG95 protein, and expression thereof in yeast | |
| CN103037888A (en) | Treatment of proliferative diseases | |
| CN103131676A (en) | Silkworm recombinant baculovirus showing recombinant human tumor necrosis factor receptor-Fc fusion protein gene, and preparing method and application thereof | |
| CN109134625B (en) | Pantoea ananatis protein exciton HCP and function thereof | |
| CN103239477A (en) | Separation and preparation method of mesenchymal stem cells, neural stem cells and active factors for stem cell induction | |
| CN102516381B (en) | Natural sex storage protein-2 as well as preparation and use for same | |
| CN107022549A (en) | Pelteobagrus fulvidraco beta-defensin gene and its beta-defensin antibacterial peptide and its application | |
| CN107540733B (en) | Rice blast bacterium exciton Mo65 and purification method thereof | |
| CN100381560C (en) | Process for synthesizing recombined human intestine trilobate factor using GS115 microzyme | |
| CN101906423A (en) | Preparation method of recombined human nerve growth factor based on insect baculovirus expression system | |
| Jockusch et al. | Early cell death caused by TMV mutants with defective coat proteins | |
| CN109161488A (en) | One plant height produces Irpex lacteus strain and its cultural method of cordycepin | |
| CN102250216B (en) | Rana nigromaculata antimicrobial peptide as well as gene and application thereof | |
| CN104342423A (en) | High activity recombinant human chymotrypsin preparation method and application thereof | |
| CN1312292A (en) | Preparation process of gene antibiotic peptide medicine from transgenic antibiotic peptide fly maggot | |
| CN106191184A (en) | A kind of preparation of novel Scapharca broughtonii antioxidation active peptides and application thereof | |
| CN106879884A (en) | A kind of method for improving royal jelly performance | |
| CN110923289A (en) | Screening method of drug for treating citrus greening disease | |
| CN104804085A (en) | Method for preparing diarrhetic shellfish poisoning okadaic acid monoclonal antibody | |
| CN104292310B (en) | Duck plague virus UL15 gene exonI recombinant proteins and its preparation method and application | |
| CN108314735A (en) | A kind of monoclonal antibody of anti-vole Babesia and its application | |
| Chakraborty et al. | Characterization of antimicrobial peptides from local forest dwelling ants: in-vitro screening for antimicrobial activity | |
| CN104491858B (en) | A kind of composition of liquid medicine containing animal wool scurf allergen of stabilization |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: TIANJIN?INTERNATIONAL?JOINT?ACADEMY?OF?BIOTECHNOLO Effective date: 20130527 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20130527 Address after: 300457 intersection of Dongting Road and thirteen Avenue, Binhai New Area Economic and Technological Development Zone, Tianjin, Tianjin International Joint Research Institute of biomedicine, S501 Applicant after: TIANJIN YAOYU BIOLOGICAL TECHNOLOGY Co.,Ltd. Applicant after: Tianjin International Joint Academy of Biotechnology & Medicine Address before: Dongting Road 300457 in Tianjin Binhai Economic and Technological Development Zone No. 220 Applicant before: TIANJIN YAOYU BIOLOGICAL TECHNOLOGY Co.,Ltd. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| PP01 | Preservation of patent right | ||
| PP01 | Preservation of patent right |
Effective date of registration: 20191202 Granted publication date: 20131030 |
|
| PD01 | Discharge of preservation of patent | ||
| PD01 | Discharge of preservation of patent |
Date of cancellation: 20221202 Granted publication date: 20131030 |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131030 |