CN101659700B - Short peptide - Google Patents
Short peptide Download PDFInfo
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- CN101659700B CN101659700B CN2009100414203A CN200910041420A CN101659700B CN 101659700 B CN101659700 B CN 101659700B CN 2009100414203 A CN2009100414203 A CN 2009100414203A CN 200910041420 A CN200910041420 A CN 200910041420A CN 101659700 B CN101659700 B CN 101659700B
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- leu
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- small peptide
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Abstract
The invention relates to a short peptide. The amino acid sequence of the small peptide is Ile Leu Ser Lys Leu Arg Leu Glu Thr Ala Pro Asn Ile Ser Lys Ala Val Ile Arg Gln Leu Leu Pro Arg Ala Pro Pro Leu Arg Glu. The invention simulates the combination mode of an M propeptide and a C segment, synthesizes the short peptide which can be combined with the C segment, can inhibit the MSTN activity in the animal bodies and promote the growth of the skeletal muscle of livestock, and is simple, convenient, safe and effective when being used for breeding the livestock with more muscles.
Description
[technical field]
The present invention relates to a kind of small peptide.
[background technology]
McPherron etc. have found a kind of negative regulatory factor that plays an important role in the muscle growth process when the research amyotrophy, result of study is after delivering on the Nature magazine, the author with this factor be named as myostatin (myostatin, MSTN).This discovery has caused correlative study personnel's very big interest, has many scholars to carry out deep research to the molecular mechanism of MSTN effect and with amyotrophy diseases associated mechanism subsequently.On clinical medicine, sports medical science and agricultural, the research of MSTN also there are many progress.According to its constructional feature, MSTN has been classified as growth and differentiation factor-8 (growth differentiation factor 8, GDF-8), (member in the superfamily of transforming growth factor-β) plays crucial down regulation in the growth of skeletal muscle to belong to TGF-β.MSTN suppresses the important cytokine that Skeletal Muscle Growth is grown, and belongs to the paracrine or the autocrine factor, secretes outside the born of the same parents after being expressed by Skeletal Muscle Cell, acts on self myocyte or Skeletal Muscle Cell on every side, suppresses myocyte's growth.Body MSTN genetic expression under standard state keeps very low level, makes muscle growth be in an equilibrium state.This transgenation or the animal that knocks out, muscle growth loses inhibition, the skeletal muscle muscle group obviously increases, show as the extraordinary prosperity of whole body skeletal muscle, as the mouse of MSTN gene knockout, skeletal muscle muscle has increased 2-3 doubly, and Belgian blue ox is because sudden change has taken place the MSTN gene, cause disinthibiting of its Skeletal Muscle Growth, shown as the improved seeds of high yield meat.And in some amyotrophic organisms, this expression of gene level then significantly raises.Body is in the state that does not move for a long time, also can cause the rise that MSTN expresses, and causes the atrophy of muscle.There is in various degree raising in the patient of many long-term beds or nutrient consumption disease as intravital MSTN levels such as AIDS VICTIMSs.Often be in the people of physical activity state, its intravital MSTN level is lower than the level in the long-term not movement human.
376 amino acid of MSTN total length, after the Skeletal Muscle Cell expression, become two fragments at the 267th amino acids place by protease cracking, the N end parts claims propeptide (myostatin propetide, be called for short the M propetide), the C end fragment is the active region of MSTN, and two C end molecules combine with non-covalent form by dimer of disulfide linkage formation and with the M propetide, and C holds in conjunction with not showing biologic activity behind the M propetide.The C fragment with enter into body fluid after the M propetide combines, subsequently by the aspartic acid place cracking of metalloprotease at the 76th, this moment, the C end just showed the biologic activity that suppresses muscle growth in conjunction with the segmental M propetide of C.This moment is if there is a large amount of M propetides to have the activity that will continue to suppress the C section.
Motion exercise can lower the expression of MSTN in the body, reduces the inhibition of MSTN to Skeletal Muscle Growth, promotes the growth of muscle then indirectly, shows as flourishing skeletal muscle.Have and experimental results show that: suppress the activity of MSTN in the body, can promote myoblastic division and breeding,, the M propetide is constantly combined with the C section, just can suppress the activity of MSTN, but used amount is bigger as giving injection M propetide in the experimental animals.Other has the investigator to adopt the antibody of anti-MSTN and external experimental research has been carried out in the effect that suppresses MSTN at the RNAi of MSTN, thinks that the two all has the activity that suppresses MSTN.But antibody belongs to heterologous protein, and use can cause transformation reactions in vivo.In the cell experiment of vitro culture, RNAi enters cell by transfection, just can't enter cell by transfection in vivo, though there are some retroviral vectors to bring RNAi into cell, but still have potential safety hazard.
[summary of the invention]
The present invention aims to provide a kind of MSTN activity that can suppress animal, and what promote Skeletal Muscle Growth has opposing " myostatin " (myostatin, MSTN) an active small peptide.
Small peptide of the present invention, its aminoacid sequence is:
Ile?Leu?Ser?Lys?Leu?Arg?Leu?Glu?Thr?Ala?Pro?Asn?Ile?Ser?Lys
Ala?Val?Ile?Arg?Gln?Leu?Leu?Pro?Arg?Ala?Pro?Pro?Leu?Arg?Glu
Perhaps be:
Ile?Leu?Ser?Lys?Leu?Arg?Leu?Glu?Thr?Ala?Pro?Asn?Ile?Ser?Lys
Ala?Ala?Ile?Arg?Gln?Leu?Leu?Pro?Lys?Ala?Pro?Pro?Leu?Arg?Glu
Perhaps be:
Pro?Leu?Thr?Val?Asp?Phe?Glu?Val?Phe?Gly?Trp?Asp?Trp?Ile?Ile
Ala?Pro?Lys?Arg?Tyr?Arg?Ala?Asn?Tyr?Cys?Ser?Gly?Glu?Cys?Glu
These small peptides have opposing ' myostatin ' (myostatin, MSTN) activity.Can be used to cultivate polymyarian meat livestock.
Small peptide of the present invention adopts known method of the prior art to carry out, and both can carry out chemosynthesis with the polypeptide automatic DNA synthesizer DNA, short peptide sequence can be derived into nucleotide sequence again, is cloned into then to carry out biosynthesizing in the expression vector.
The present invention simulates M propetide and C section bonded pattern, synthetic can with C section bonded small peptide, suppress the intravital MSTN activity of animal, can promote the livestock Skeletal Muscle Growth, when being used to cultivate polymyarian meat livestock, simple, convenient, safety, effectively.
[embodiment]
Embodiment 1 preparation small peptide of the present invention
As peptide synthetic solid-phase matrix,, hold beginning to carry out successively from C with resin according to the aminoacid sequence of small peptide of the present invention according to the solid state chemistry synthetic method of Merrifield invention.In the building-up reactions post, carry out reaction solution decompressing and extracting, wash steps such as resin.Wash resin process successively respectively with Virahol, methylene dichloride, dimethyl formamide, contain the dimethyl formamide deprotection solution reaction of 6-pyridinium hydroxide, drain reaction column.Subsequently the synthetic peptide on the resin is carried out cracking, lysate adopts trifluoroacetic acid, methyl-phenoxide, dithioglycol to carry out under the lucifuge condition, is dissolved in the cold diethyl ether at last.Obtain rough peptide product after the lyophilize.The raw product of peptide is trifluoroacetic acid and acetonitrile with the reverse column separating purification of C18, the moving phase of sepn process, through obtaining the pure product of peptide of the present invention behind the gradient elution.
Among the following embodiment, the aminoacid sequence of described small peptide 1 is:
Ile?Leu?Ser?Lys?Leu?Arg?Leu?Glu?Thr?Ala?Pro?Asn?Ile?Ser?Lys
Ala?Val?Ile?Arg?Gln?Leu?Leu?Pro?Arg?Ala?Pro?Pro?Leu?Arg?Glu
The aminoacid sequence of small peptide 2 is:
Ile?Leu?Ser?Lys?Leu?Arg?Leu?Glu?Thr?Ala?Pro?Asn?Ile?Ser?Lys
Ala?Ala?Ile?Arg?Gln?Leu?Leu?Pro?Lys?Ala?Pro?Pro?Leu?Arg?Glu
The aminoacid sequence of small peptide 3 is:
Pro?Leu?Thr?Val?Asp?Phe?Glu?Val?Phe?Gly?Trp?Asp?Trp?Ile?Ile
Ala?Pro?Lys?Arg?Tyr?Arg?Ala?Asn?Tyr?Cys?Ser?Gly?Glu?Cys?Glu
Embodiment 2 small peptides promote the experiment of small white mouse Skeletal Muscle Growth
20 of common male Kunming kind small white mouses, body weight is limited to 23g ± 3g.Mouse is divided into 4 groups, 1 group of small peptide, and 2 groups of small peptides, 3 groups of small peptides, other establishes one group of control group of not injecting small peptide.Every group 5, every mouse peritoneal was injected small peptide 1mg/ days, and injection is every other day once injected 15 times altogether.Come respectively to organize the effect of small peptide with the relative weight in wet base of ' hind leg soleus muscle/mouse body weight ' at last to Skeletal Muscle Growth.The method of calculation of weight in wet base are relatively: soleus muscle weight (mg) and the ratio of mouse from heavy sensation of the whole body (g).
As a result, the relative weight in wet base that small peptide is 1 group is 3.2mg/g ± 0.6, and the relative weight in wet base that small peptide is 2 groups is 3.5mg/g ± 0.3, and the relative weight in wet base that small peptide is 3 groups is 3.6mg/g ± 0.5, and the relative weight in wet base of control group is 2.4mg/g ± 0.5.
The result shows the relative weight in wet base of small peptide group of the present invention apparently higher than control group, illustrates that small peptide of the present invention (small peptide 1, small peptide 2, small peptide 3) has the effect of tangible promotion Skeletal Muscle Growth to small white mouse.
Embodiment 3 small peptides promote the experiment of pig Skeletal Muscle Growth
Big young pig was 12 in one month, was divided into 4 groups, 3 every group.1 group of every small peptide, another group is not for injecting the control group of small peptide.Every young pig per injection small peptide 10mg, injection every other day once.Relative weight in wet base of deltoid muscle of relatively respectively organizing after 30 days.
As a result, the relative weight in wet base of 1 group of small peptide, 2 groups of small peptides, 3 groups of small peptides has improved 30%, 19%, 12% respectively than the relative weight in wet base of control group pig deltoid muscle.Illustrate that small peptide of the present invention (small peptide 1, small peptide 2, small peptide 3) has the effect of tangible promotion pig Skeletal Muscle Growth.
Embodiment 4 small peptides promote the experiment of ox Skeletal Muscle Growth
Big calf was 8 in two months, was divided into 4 groups, 2 every group.1 group of every small peptide, another group is not for injecting the control group of small peptide.Every calf per injection small peptide 50mg, injection every other day once.Respectively organize the relative weight in wet base of deltoid muscle after 45 days,
As a result, the relative weight in wet base of 1 group of small peptide, 2 groups of small peptides, 3 groups of small peptides has improved 25%, 20%, 16% respectively than the relative weight in wet base of control calves deltoid muscle.Illustrate that small peptide of the present invention (small peptide 1, small peptide 2, small peptide 3) has the effect of tangible promotion calf Skeletal Muscle Growth.
Embodiment 5 small peptides promote the experiment of goat Skeletal Muscle Growth
Big goat was 12 in one month, was divided into 4 groups, 3 every group.1 group of every small peptide, another group is not for injecting the control group of small peptide.Every goat per injection small peptide 10mg, injection every other day once.Respectively organize the relative weight in wet base of deltoid muscle after 45 days.
As a result, the relative weight in wet base of 1 group of small peptide, 2 groups of small peptides, 3 groups of small peptides has improved 28%, 22%, 18% respectively than the relative weight in wet base of control group goat deltoid muscle.Illustrate that small peptide of the present invention (small peptide 1, small peptide 2, small peptide 3) has the effect of tangible promotion goat Skeletal Muscle Growth.
Sequence table
＜110〉Guangzhou Rosson Bioscience Co., Ltd.
＜120〉a kind of small peptide
<160>3
<210>1
<211>30
<212>PRT
＜213〉artificial sequence
<400>1
Ile?Leu?Ser?Lys?Leu?Arg?Leu?Glu?Thr?Ala?Pro?Asn?Ile?Ser?Lys
1 5 10 15
Ala?Val?Ile?Arg?Gln?Leu?Leu?Pro?Arg?Ala?Pro?Pro?Leu?Arg?Glu
20 25 30
<210>2
<211>30
<212>PRT
＜213〉artificial sequence
<400>2
Ile?Leu?Ser?Lys?Leu?Arg?Leu?Glu?Thr?Ala?Pro?Asn?Ile?Ser?Lys
1 5 10 15
A1a?Ala?Ile?Arg?Gln?Leu?Leu?Pro?Lys?Ala?Pro?Pro?Leu?Arg?Glu
20 25 30
<210>3
<211>30
<212>PRT
＜213〉artificial sequence
<400>3
Pro?Leu?Thr?Val?Asp?Phe?Glu?Val?Phe?Gly?Trp?Asp?Trp?Ile?Ile
1 5 10 15
Ala?Pro?Lys?Arg?Tyr?Arg?Ala?Asn?Tyr?Cys?Ser?Gly?Glu?Cys?Glu
20 25 30
Claims (3)
1. small peptide is characterized in that its aminoacid sequence is:
Ile?Leu?Ser?Lys?Leu?Arg?Leu?Glu?Thr?Ala?Pro?Asn?Ile?Ser?Lys
Ala?Val?Ile?Arg?Gln?Leu?Leu?Pro?Arg?Ala?Pro?Pro?Leu?Arg?Glu
2. small peptide is characterized in that its aminoacid sequence is:
Ile?Leu?Ser?Lys?Leu?Arg?Leu?Glu?Thr?Ala?Pro?Asn?Ile?Ser?Lys
Ala?Ala?Ile?Arg?Gln?Leu?Leu?Pro?Lys?Ala?Pro?Pro?Leu?Arg?Glu
3. the application of small peptide as claimed in claim 1 or 2 in cultivating polymyarian meat livestock.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100414203A CN101659700B (en) | 2009-07-27 | 2009-07-27 | Short peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100414203A CN101659700B (en) | 2009-07-27 | 2009-07-27 | Short peptide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101659700A CN101659700A (en) | 2010-03-03 |
| CN101659700B true CN101659700B (en) | 2011-11-16 |
Family
ID=41787972
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100414203A Expired - Fee Related CN101659700B (en) | 2009-07-27 | 2009-07-27 | Short peptide |
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| Country | Link |
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| CN (1) | CN101659700B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103483049B (en) * | 2012-10-30 | 2014-11-12 | 山东红日阿康化工股份有限公司 | Heavy metal remover for water soluble fertilizer |
| CN110935006B (en) * | 2019-11-12 | 2021-05-18 | 中食都庆（山东）生物技术有限公司 | Application of earthworm protein peptide in preparing medicine for preventing and/or treating thrombotic diseases |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1514020A (en) * | 2003-05-30 | 2004-07-21 | 哈尔滨工业大学 | Mutant chicken myostatin and detecting method of said gene and application in chicken breeding |
| CN1569230A (en) * | 2003-11-05 | 2005-01-26 | 华南农业大学 | Preparation and application of a gene engineering vaccine of muscle growth retarder |
| WO2008016314A1 (en) * | 2006-08-03 | 2008-02-07 | Orico Limited | Myostatin antagonists |
-
2009
- 2009-07-27 CN CN2009100414203A patent/CN101659700B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1514020A (en) * | 2003-05-30 | 2004-07-21 | 哈尔滨工业大学 | Mutant chicken myostatin and detecting method of said gene and application in chicken breeding |
| CN1569230A (en) * | 2003-11-05 | 2005-01-26 | 华南农业大学 | Preparation and application of a gene engineering vaccine of muscle growth retarder |
| WO2008016314A1 (en) * | 2006-08-03 | 2008-02-07 | Orico Limited | Myostatin antagonists |
Non-Patent Citations (1)
| Title |
|---|
| 刘超武等.逆转录病毒载体介导的RNA干扰稳定抑制肌肉生长抑制素GDF-8的表达.《生物工程学报》.2008,第2卷(第02期),250-255. * |
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| Publication number | Publication date |
|---|---|
| CN101659700A (en) | 2010-03-03 |
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Owner name: HUNAN PROVINCE OF ANIMAL HUSBANDRY INDUSTRY LIMITE Free format text: FORMER OWNER: GUANGZHOU ROSSON BIOSCIENCE CO., LTD. Effective date: 20130106 |
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Free format text: CORRECT: ADDRESS; FROM: 510663 GUANGZHOU, GUANGDONG PROVINCE TO: 422800 SHAOYANG, HUNAN PROVINCE |
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Effective date of registration: 20130106 Address after: 422800 Shaodong County of Hunan province Niumasi town wells village 1 groups Patentee after: Animal husbandry in Hunan Province Asia Pharmaceutical Co., Ltd. Address before: 510663 Guangdong province Guangzhou Science City skim Springs Road No. 3, international business incubator building D room 908 Patentee before: Guangzhou Rosson Bioscience Co., Ltd. |
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