JPH0446197A - New protein - Google Patents
New proteinInfo
- Publication number
- JPH0446197A JPH0446197A JP15409990A JP15409990A JPH0446197A JP H0446197 A JPH0446197 A JP H0446197A JP 15409990 A JP15409990 A JP 15409990A JP 15409990 A JP15409990 A JP 15409990A JP H0446197 A JPH0446197 A JP H0446197A
- Authority
- JP
- Japan
- Prior art keywords
- amino acid
- protein
- molecular weight
- gly
- collected
- 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
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 22
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 21
- 125000000539 amino acid group Chemical group 0.000 claims abstract description 7
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 claims abstract description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 241000555745 Sciuridae Species 0.000 abstract description 8
- 239000008280 blood Substances 0.000 abstract description 6
- 210000004369 blood Anatomy 0.000 abstract description 6
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 3
- 230000015271 coagulation Effects 0.000 abstract description 3
- 238000005345 coagulation Methods 0.000 abstract description 3
- 238000002523 gelfiltration Methods 0.000 abstract description 3
- 229960002897 heparin Drugs 0.000 abstract description 3
- 229920000669 heparin Polymers 0.000 abstract description 3
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 3
- 238000001502 gel electrophoresis Methods 0.000 abstract description 2
- 229920002401 polyacrylamide Polymers 0.000 abstract description 2
- 230000005855 radiation Effects 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 230000006266 hibernation Effects 0.000 description 9
- 150000001413 amino acids Chemical group 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 108020004635 Complementary DNA Proteins 0.000 description 3
- 238000010804 cDNA synthesis Methods 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 230000007958 sleep Effects 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- NKJBKNVQHBZUIX-ACZMJKKPSA-N Ala-Gln-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O NKJBKNVQHBZUIX-ACZMJKKPSA-N 0.000 description 1
- DPNZTBKGAUAZQU-DLOVCJGASA-N Ala-Leu-His Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N DPNZTBKGAUAZQU-DLOVCJGASA-N 0.000 description 1
- NONSEUUPKITYQT-BQBZGAKWSA-N Arg-Asn-Gly Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N)CN=C(N)N NONSEUUPKITYQT-BQBZGAKWSA-N 0.000 description 1
- VTYQAQFKMQTKQD-ACZMJKKPSA-N Asp-Ala-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(O)=O VTYQAQFKMQTKQD-ACZMJKKPSA-N 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 108091028026 C-DNA Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 241000208296 Datura Species 0.000 description 1
- 241001416537 Gliridae Species 0.000 description 1
- LKDIBBOKUAASNP-FXQIFTODSA-N Glu-Ala-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LKDIBBOKUAASNP-FXQIFTODSA-N 0.000 description 1
- NCWOMXABNYEPLY-NRPADANISA-N Glu-Ala-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O NCWOMXABNYEPLY-NRPADANISA-N 0.000 description 1
- NLKVNZUFDPWPNL-YUMQZZPRSA-N Glu-Arg-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O NLKVNZUFDPWPNL-YUMQZZPRSA-N 0.000 description 1
- RJONUNZIMUXUOI-GUBZILKMSA-N Glu-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)O)N RJONUNZIMUXUOI-GUBZILKMSA-N 0.000 description 1
- SJLKKOZFHSJJAW-YUMQZZPRSA-N Gly-Met-Glu Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)CN SJLKKOZFHSJJAW-YUMQZZPRSA-N 0.000 description 1
- JBCLFWXMTIKCCB-VIFPVBQESA-N Gly-Phe Chemical compound NCC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-VIFPVBQESA-N 0.000 description 1
- JPVGHHQGKPQYIL-KBPBESRZSA-N Gly-Phe-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 JPVGHHQGKPQYIL-KBPBESRZSA-N 0.000 description 1
- UVTSZKIATYSKIR-RYUDHWBXSA-N Gly-Tyr-Glu Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O UVTSZKIATYSKIR-RYUDHWBXSA-N 0.000 description 1
- BNMRSWQOHIQTFL-JSGCOSHPSA-N Gly-Val-Phe Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 BNMRSWQOHIQTFL-JSGCOSHPSA-N 0.000 description 1
- 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 1
- RLAOTFTXBFQJDV-KKUMJFAQSA-N His-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CN=CN1 RLAOTFTXBFQJDV-KKUMJFAQSA-N 0.000 description 1
- YKLOMBNBQUTJDT-HVTMNAMFSA-N Ile-His-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N YKLOMBNBQUTJDT-HVTMNAMFSA-N 0.000 description 1
- AKVBOOKXVAMKSS-GUBZILKMSA-N Leu-Ser-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O AKVBOOKXVAMKSS-GUBZILKMSA-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
- KQBJYJXPZBNEIK-DCAQKATOSA-N Met-Glu-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQBJYJXPZBNEIK-DCAQKATOSA-N 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- SEPNOAFMZLLCEW-UBHSHLNASA-N Phe-Ala-Val Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)O SEPNOAFMZLLCEW-UBHSHLNASA-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
- CGBYDGAJHSOGFQ-LPEHRKFASA-N Pro-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2 CGBYDGAJHSOGFQ-LPEHRKFASA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- BQWCDDAISCPDQV-XHNCKOQMSA-N Ser-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CO)N)C(=O)O BQWCDDAISCPDQV-XHNCKOQMSA-N 0.000 description 1
- SFTZWNJFZYOLBD-ZDLURKLDSA-N Ser-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO SFTZWNJFZYOLBD-ZDLURKLDSA-N 0.000 description 1
- DKDHTRVDOUZZTP-IFFSRLJSSA-N Thr-Gln-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O DKDHTRVDOUZZTP-IFFSRLJSSA-N 0.000 description 1
- CKHWEVXPLJBEOZ-VQVTYTSYSA-N Thr-Val Chemical compound CC(C)[C@@H](C([O-])=O)NC(=O)[C@@H]([NH3+])[C@@H](C)O CKHWEVXPLJBEOZ-VQVTYTSYSA-N 0.000 description 1
- LYMVXFSTACVOLP-ZFWWWQNUSA-N Trp-Leu Chemical compound C1=CC=C2C(C[C@H]([NH3+])C(=O)N[C@@H](CC(C)C)C([O-])=O)=CNC2=C1 LYMVXFSTACVOLP-ZFWWWQNUSA-N 0.000 description 1
- DCOOGDCRFXXQNW-ZKWXMUAHSA-N Val-Asn-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CS)C(=O)O)N DCOOGDCRFXXQNW-ZKWXMUAHSA-N 0.000 description 1
- VPGCVZRRBYOGCD-AVGNSLFASA-N Val-Lys-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O VPGCVZRRBYOGCD-AVGNSLFASA-N 0.000 description 1
- KISFXYYRKKNLOP-IHRRRGAJSA-N Val-Phe-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)O)N KISFXYYRKKNLOP-IHRRRGAJSA-N 0.000 description 1
- SJRUJQFQVLMZFW-WPRPVWTQSA-N Val-Pro-Gly Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SJRUJQFQVLMZFW-WPRPVWTQSA-N 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010020688 glycylhistidine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 230000004217 heart function Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 108010080629 tryptophan-leucine Proteins 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は新規蛋白質に間するものである０番くは、動物
の冬眠現象と密接な関連を有する新短な蛋白質に間する
ものである。[Detailed Description of the Invention] (Industrial Application Field) The present invention relates to a novel protein. .
（発明が解決しようとする課１！ＩＮ）シマリス、ヤマ
ネ等の鴫乳動物が、冬朋に活動性を失い水や食餌を殆ん
と摂取せずに生命を維持する、所謂冬眠現象については
古くから知られているが、その実態については充分解明
されていない０本発明は冬眠現象の発現を左右する因子
を解明することを目的とするものである。(Question 1 to be solved by the invention!IN) Regarding the so-called hibernation phenomenon, in which mammals such as chipmunks and dormice lose their activity during the winter and maintain their lives without ingesting much water or food. Although it has been known for a long time, the actual situation has not been fully elucidated.The purpose of the present invention is to elucidate the factors that influence the manifestation of the hibernation phenomenon.
（課題を解決するための手段）
本発明者等は、さきにシマリスについて長期間の低温暴
露による冬眠誘導実験を行ない、冬眠時の心臓機能につ
いて検討を行ったところ、冬眠下においては、心臓の電
気的機械的性質が著しく変化しており、この変化は冬眠
開始前から始っていることが判明した。(Means for Solving the Problems) The present inventors previously conducted a hibernation induction experiment on chipmunks by exposing them to low temperatures for a long period of time, and investigated cardiac function during hibernation. It was found that the electrical and mechanical properties of the animals changed significantly, and that these changes began before the onset of hibernation.
上記の事実に着目して検討を進めた結果、非冬眠状態の
シマリスの血液中に存在するある種の蛋白質が、冬眠の
開始前から徐々に減少し、冬眠時には殆んど消失してい
ることを確認した。本発明は上記の知見に基づいて更に
検討を重ねた結果、この蛋白質を単離し、性状及び構造
を解明することにより本発明を達成した。As a result of conducting studies focusing on the above facts, we found that a certain type of protein present in the blood of non-hibernating chipmunks gradually decreases before the start of hibernation, and almost disappears during hibernation. It was confirmed. As a result of further studies based on the above findings, the present invention was achieved by isolating this protein and elucidating its properties and structure.
即ち、本発明の要旨は、ドデシル硫酸ナトリウム−ポリ
アクリルアミドゲル電気泳動の検定による分子量が約２
７キロダルトンであり、下記のアミノ酸配列を有し、か
つそのＮ末端の約４０のアミノ酸残基中にＧｌｙ−Ｘ−
Ｙ（Ｘ及びＹは任意のアミノ酸残基を示す）の繰り返し
配列をもつコラーゲン様構造を有する新規蛋白質に存す
る。That is, the gist of the present invention is that the molecular weight as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is approximately 2.
7 kilodaltons, has the following amino acid sequence, and has Gly-X-
It is a novel protein with a collagen-like structure having a repeating sequence of Y (X and Y represent arbitrary amino acid residues).
工上工Ｖａｌ Ａｓｎ Ｃｙｓ Ｈｊｓ Ｓｅｒ Ｌ
ｙｓ Ｇｌｙ ’Ｔｈｒ ５ｅｒＡｌａ Ｐｈｅ
Ａｌａ Ｖａｌ Ｌｙｓ Ａｌａ Ａｓｎ Ｇｌｕ
Ｌｅｕ Ｐｒ。Construction Val Asn Cys Hjs Ser L
ys Gly 'Thr 5erAla Phe
Ala Val Lys Ala Asn Glu
Leu Pr.
Ｐｒｏ Ａｌａ Ｐｒｏ Ｓｅｒ Ｇｌｎ Ｐｒｏ
Ｖａｌ Ｔ Ｉｅ Ｐｈｅ ＬｙｓＧｌｕ Ａｌａ
Ｌｅｕ Ｈｉｓ Ａｓｐ Ａｌａ Ｇｌｎ Ｇｌｙ
Ｈｉｓ ＰｈｅＡｓｐ Ｌｅｕ Ａｌａ Ｔｂｒ
Ｇｌｙ Ｖａｌ Ｐｈｅ Ｔｈｒ Ｃｙｓ
Ｐｒ。Pro Ala Pro Ser Gln Pro
Val T Ie Phe LysGlu Ala
Leu His Asp Ala Gln Gly
His PheAsp Leu Ala Tbr
Gly Val Phe Thr Cys
Pr.
Ｖａｌ Ｐｒｏ Ｇｌｙ Ｌｅｕ Ｔｙｒ Ｇ
ｉｎ Ｐｂｅ Ｇｌｙ Ｐｈｅ ＨｌｓＩ Ｉｅ
Ｇｌｕ Ａｌａ Ｖａｌ Ｇｌｎ Ａｒｇ Ａｌ
ａ Ｖａｌ Ｌｙｓ ＶａｌＳｅｒ Ｌｅｕ Ｍ
ｅｔ Ａｒｇ Ａｓｎ Ｇｌｙ Ｔｈｒ Ｇｌｎ
Ｖａｌ ＭｅｔＧｌｕ Ａｒｇ Ｇｌｕ Ａｌａ
Ｇｌｕ Ａｌａ Ｇｌｎ Ａｓｐ Ｇｌｙ Ｔｙｒ
Ｇｌｕ Ｈｉｓ ｒ Ｉｅ Ｓｅｒ Ｇｌｙ
Ｔｈｒ Ａｌａ Ｔｉｅ Ｌｅｕ ＧｉｎＬｅｕ
Ｇｌｙ Ｍｅｔ Ｇｌｕ Ａｓｐ Ａｒｇ Ｖａ
ｔ Ｔｒｐ Ｌｅｕ ＧｌｕＡｓｎ Ｌｙｓ
Ｌｅｕ Ｓｅｒ Ｇｌｎ Ｔｈｒ Ａｓｐ Ｌｅ
ｕ Ｇｌｕ ＡｒｇＧｌｙ Ｔｈｒ Ｖａｌ Ｇ
ｌｎ Ａｌａ Ｖａｌ Ｐｈｅ Ｓｅｒ Ｇｌｙ
ＰｈｅＬｅｕ Ｉｌｅ Ｈｉｓ Ｇｌｕ Ａ
ｓｎ以下に本発明の詳細な説明する。Val Pro Gly Leu Tyr G
in Pbe Gly Phe HlsI Ie
Glu Ala Val Gln Arg Al
a Val Lys Val Ser Leu M
et Arg Asn Gly Thr Gln
Val MetGlu Arg Glu Ala
Glu Ala Gln Asp Gly Tyr
Glu His r Ie Ser Gly
Thr Ala Tie Leu GinLeu
Gly Met Glu Asp Arg Va
t Trp Leu GluAsn Lys
Leu Ser Gln Thr Asp Le
u Glu ArgGly Thr Val G
ln Ala Val Phe Ser Gly
PheLeu Ile His Glu A
The present invention will be described in detail below.
本発明の蛋白質は、次の方法により単層することができ
る。即ち、シマリスの血液を採取し、ヘパリンを添加し
て凝固を阻止した後、遠心分離して血漿を採取する。得
られた血漿を高速液体クロマトグラフィー（ＨＰＬＣ）
によりゲル濾過カラムを用いて分画し、約１４０キロダ
ルトン（ｋＤ）の分子量に相当する蛋白質画分を採取す
る。The protein of the present invention can be formed into a monolayer by the following method. That is, the chipmunk's blood is collected, heparin is added to prevent coagulation, and the blood is centrifuged to collect plasma. The obtained plasma was subjected to high performance liquid chromatography (HPLC).
The protein fraction is fractionated using a gel filtration column, and a protein fraction corresponding to a molecular weight of about 140 kilodaltons (kD) is collected.
この両分を濃縮、脱塩して得られた濃縮液から、ドデシ
ル硫酸ナトリウム（ＳＯ５）−ポリアクリルアミドゲル
のスラブゲル電気泳動により、単一バントを示す分子置
駒２７ｋＤの本発明の蛋白質が得られる。From the concentrated solution obtained by concentrating and desalting both components, the protein of the present invention having a molecular weight of 27 kD and exhibiting a single band can be obtained by slab gel electrophoresis on sodium dodecyl sulfate (SO5)-polyacrylamide gel. .
こうして単離された蛋白質は、シマリスの肝臓から単離
したｍ −ＲＮ Ａを用いて常法によりクロニングした
相補ＤＮＡ（ｃ−ＤＮＡ）の塩基配列から、前記のアミ
ノ酸配列を有する新規な構造のものであることが判明し
た。また、そのＮ末端の約４０のアミノ酸残基中には、
前記アミノ酸配列中の下線部分に示すように、Ｇｌｙ−
Ｘ−Ｙ（Ｘ及びＹは任意のアミノ酸残基を示す）の繰り
返し配列が存在し、結合組織の基質の主成分であるコラ
ーゲン擾の構造を含有していることが判明した。The protein thus isolated has a novel structure having the above-mentioned amino acid sequence based on the base sequence of complementary DNA (c-DNA) cloned using m-RNA isolated from the liver of a chipmunk by a conventional method. It turned out to be. In addition, among the approximately 40 amino acid residues at the N-terminus,
As shown in the underlined portion of the amino acid sequence, Gly-
It was found that a repeating sequence of X-Y (X and Y represent arbitrary amino acid residues) exists, and contains the structure of collagen membranes, which are the main components of connective tissue matrix.
（発明の効果）
哺乳動物の冬眠の誘導は睡眠に引き続いて起ることから
、睡眠１１能が拡張されたものと考えられ、本発明の蛋
白質は睡眠機構の制御に関与することが推測される。ま
た冬眠中は細菌感染や、放射能、低温等による障害を受
は難いことから、この蛋白質は、これらに対する身体の
抵抗性の調節に寄与することが期待される。(Effects of the Invention) Since the induction of hibernation in mammals occurs following sleep, it is thought that the sleep function 11 is expanded, and it is speculated that the protein of the present invention is involved in the control of the sleep mechanism. . Additionally, since animals are less susceptible to damage from bacterial infections, radiation, low temperatures, etc. during hibernation, this protein is expected to contribute to regulating the body's resistance to these.
（実施例）
以下本発明を実施例について更に詳細に説明するが、本
発明はその要旨を超えない限り本実施例に限定されるも
のではない。(Example) The present invention will be described in more detail with reference to Examples below, but the present invention is not limited to the Examples unless it exceeds the gist thereof.
実施例１
チョウセンシマリスの血液を採取し、これにヘパリンを
添加して凝固を阻止した後、遠心分離し・て血漿を採取
した。得られた血漿６０μｍを、リン酸緩衝生理液（ｐ
Ｈ６，９５）を用い、高速液体クロマトグラフィー（Ｒ
ＰＬＣ）により流速０．５ ｍｌ／分でゲル濾過（カラ
ム：Ａｓａｈｉｐａｋ Ｇ５−５２０）を行ない、ＩＩ
ＰＬｃ用分子量指標蛋白（オリエンタル酵母工業社製）
により決定される約＋４０ ｋＤの分子量に相当する蛋
白質画分１−１を得た。Example 1 Blood from a Datura chipmunk was collected, heparin was added to it to prevent coagulation, and the blood was centrifuged to collect plasma. 60 μm of the obtained plasma was diluted with phosphate buffered physiological solution (p
H6,95) using high performance liquid chromatography (R
Gel filtration (column: Asahipak G5-520) was performed at a flow rate of 0.5 ml/min using II
Molecular weight indicator protein for PLc (manufactured by Oriental Yeast Industries)
Protein fraction 1-1 was obtained, corresponding to a molecular weight of approximately +40 kD as determined by .
次いで、この両分をセントリコン−１０（ブレース社！
１）を用い、濃縮、脱塩して６０μｍの濃縮液を得た。Next, these two parts were purchased from Sentricon-10 (Brace Inc.).
1) was used to concentrate and desalt to obtain a 60 μm concentrated solution.
この濃縮液３０μｍから、５ＤＳ−ポリアクリルアミド
ゲル（アクリルアミド１２．５％含有）のスラブゲル電
気泳動（３０ｔａ＾、１．５特賞）により、蛋白質の染
色液（Ｃｏｏｍａｓｓｉｅ ｂｒｉｌｌｉａｎｔ ｂｌ
ｕｅ）で染色される分子置駒２７ｋＯの本発明の蛋白質
を単一バントとして得た。上述の方法によって単離した
蛋白質は、シマリスの肝臓から単離したｍ −ＲＮ Ａ
を用い、常法によりクローニングした相補ＤＮＡ（ｃ
−Ｄ Ｎ Ａ、 ）の塩基配列から、前記アミノ酸配列
を有する新規な構造のものであることが判明した。From this concentrated solution of 30 μm, a protein staining solution (Coomassie brilliant BL
A protein of the present invention with a molecular weight of 27 kO stained with UE) was obtained as a single bundle. The protein isolated by the above method was m-RNA isolated from chipmunk liver.
Complementary DNA (c
-DNA, ) was found to have a novel structure having the above amino acid sequence.
Claims (1)
ル電気泳動の検定による分子量が約２７キロダルトンで
あり、下記のアミノ酸配列を有し、かつそのＮ末端の約
４０のアミノ酸残基中にＧｌｙ−Ｘ−Ｙ（Ｘ及びＹは任
意のアミノ酸残基を示す）の繰り返し配列をもつコラー
ゲン様構造を有する新規蛋白質。 ［Ｎ末端］ 【遺伝子配列があります】(1) It has a molecular weight of about 27 kilodaltons as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, has the following amino acid sequence, and has Gly-X-Y in about 40 amino acid residues at its N-terminus. A novel protein with a collagen-like structure having a repeating sequence of (X and Y represent arbitrary amino acid residues). [N-terminus] [Gene sequence available]
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15409990A JP2863270B2 (en) | 1990-06-14 | 1990-06-14 | Novel protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15409990A JP2863270B2 (en) | 1990-06-14 | 1990-06-14 | Novel protein |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0446197A true JPH0446197A (en) | 1992-02-17 |
| JP2863270B2 JP2863270B2 (en) | 1999-03-03 |
Family
ID=15576897
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP15409990A Expired - Lifetime JP2863270B2 (en) | 1990-06-14 | 1990-06-14 | Novel protein |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2863270B2 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007139708A (en) * | 2005-11-22 | 2007-06-07 | Toyo Tire & Rubber Co Ltd | Tire abrasion test method |
-
1990
- 1990-06-14 JP JP15409990A patent/JP2863270B2/en not_active Expired - Lifetime
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007139708A (en) * | 2005-11-22 | 2007-06-07 | Toyo Tire & Rubber Co Ltd | Tire abrasion test method |
| US7819000B2 (en) | 2005-11-22 | 2010-10-26 | Toyo Tire & Rubber Co., Ltd. | Tire wear test method |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2863270B2 (en) | 1999-03-03 |
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