蕲蛇蛇毒中一种新型金属蛋白酶AA-MP-I的纯化和表征

天然产物研究与开发 ›› 2010, Vol. 23 ›› Issue (6): 929-933.

• 研究论文 • 下一篇

蕲蛇蛇毒中一种新型金属蛋白酶AA-MP-I的纯化和表征

潘剑茹¹，何火聪²，刘枋²，饶平凡^1*

¹福州大学生物工程研究所，福州 350002；²福建省肿瘤医院，福州 350014

收稿日期:2009-03-24 出版日期:2011-01-10 发布日期:2011-10-19
通讯作者: pfrao@fzu.edu.cn
基金资助:
福建省科技厅重点项目（2000Z143）

Purification and Characterization of a Novel Metalloproteinase AA-MP-I from the Venom of Agkistrodon acutus

PAN Jian-ru¹, HE Huo-cong², LIU Fang², RAO Ping-fan^1*

¹Institute of Biotechnology, Fuzhou University, Fuzhou 350002, China; ²Fujian Provincial Tumor Hospital, Fuzhou 350014, China

Received:2009-03-24 Online:2011-01-10 Published:2011-10-19

摘要/Abstract

摘要：

本研究通过嗜硫色谱、Sephadex G-75、蓝胶和POROS HQ 20离子交换色谱，从蕲蛇蛇毒中分离得到一种新组分AA-MP-I。该酶为分子量22.9 kDa的单体蛋白，等电点为5.55，不含中性糖基，N端序列为STEFQRYMEIVIVVDHSMVK，结果表明其为新型P-I型金属蛋白酶，对温度敏感，具有抗凝血活性，40 ℃下抗凝血活性最强，具有出血毒性，无磷脂酶A₂活性。

关键词: 蕲蛇, 蛇毒, 金属蛋白酶, 抗凝血, 出血毒

Abstract:

A novel P-I snake venom metalloproteinase AA-MP-I was purified from the venom of Agkistrodon acutus by using thiophilic adsorption chromatography, Sephadex G-75, Affi-gel blue gel, and POROS HQ 20 anion-exchange chromatography. AA-MP-I was a monomer with the molecular weight of approximate 22.9 kDa on SDS-PAGE and its isoelectric point was 5.55 analyzed by IEF. AA-MP-I hasn’t any neutral carbohydrate and its N-terminal amino acid sequence was STEFQRYMEIVIVVDHSMVK. AA-MP-I has basic proteolytic and antithrombin activities, both of which are heat-unstable. In addition, AA-MP-I was proved to have hemorrhagic activity but no acidic phospholipase A₂ activity.

Key words: Agkistrodon acutus, venom, metalloproteinase, antithrombin, hemorrhage activity

潘剑茹，何火聪，刘枋，饶平凡. 蕲蛇蛇毒中一种新型金属蛋白酶AA-MP-I的纯化和表征[J]. 天然产物研究与开发, 2010, 23(6): 929-933.

PAN Jian-Ru, HE Huo-Cong, LIU Fang, RAO Ping-Fan. Purification and Characterization of a Novel Metalloproteinase AA-MP-I from the Venom of Agkistrodon acutus[J]. NATURAL PRODUCT RESEARCH AND DEVELOPMENT, 2010, 23(6): 929-933.

参考文献

1 Xue WM(薛伟明),Guo CT(郭春腾),Chen TB(陈天豹),et al.Purification and characterization of a thrombin-like enzyme from Agkistrodon acutus.J Fuzhou Univ, Nat Sci(福州大学学报,自科版),2001, 29:112-115.
2 Lai WP(赖伟苹),Guo CT(郭春腾),Den WH(邓文汉),et al.Purification and characterization of a novel thrombin-like enzyme from Agkistrodon acutus Venom.Life Sci Res(生命科学研究),2002,6:64-67.
3 Xu ZB(许志宾),Guo CT(郭春腾),Wang ZL(王中来),et al.Purification and characterization of an anticoagulant factor from the venom of Agkistuodon acutus.Journal of Snake(蛇志),2003,15:21-24.
4 Rawlings ND,Barrett AJ.Evolutionary families of metallopeptidases.Methods Enzymol,1995,248: 183-228.
5 Takeya H,Oda K,Miyata T,et al.The complete amino acid sequence of the high molecular mass hemorrhagic protein HR1B isolated from the venom of Trimeresurus flavoviridis.J Biol Chem,1990. 265:16068-16073.
6 Hite LA,Jia LJ,Bjarnason JB,et al.cDNA sequences for four snake venom metalloproteinases:structure, classification,and their relationship to mammalian reproductive proteins.Arch Biochem Biophys,1994.308: 182-191.
7 Xu X,Wang C,Liu J,et al.Purification and characterization of hemorrhagic components from Agkistrodon acutus (hundred pace snake) venom.Toxicon,1981,19:633-644.
8 Nikai T,Kato C,Komori Y,et al.Primary structure of Ac-1 proteinase from the venom of Deinagkistrodon acutus (Hundred-pace snake) from Taiwan.Biol Pharm Bull,1995,18:631-633.
9 Liu QC,Xu HX,Cheng X,et al.Molecular cloning and sequence analysis of cDNA hemonhagic toxin acutolysin a fram agkistrodon acutus.Toxicon,1999,37:1539-1548.
10 Liu QD(刘庆都),Xu WH(徐卫华),Cheng X(承新),et al.Molecular cloning and sequence analysis of cDNA encoding acutolysin C,a Hemorrhagic metalloproteinase, from Agkistrodon acutus.Acta Biochimica et Biophysica Sinica(生物化学与生物物理学报) 2000,32:425-429 .
11 Scoble JA,Scopes RK.Ligand structure of the divinylsulfone-based T-gel.J Chromatogr A,1997,787: 47-54.
12 Laemmli UK.Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature,1970,227,(5259):680-685.
13 Zhao YF(赵永芳).Technique Principle and Application Of Biochemistry (生物化学技术原理及应用). Beijing:Science Press,2002.395-398.
14 Dubois M,Gilles KA, Hamilton JK,et al.Colorimetric method for determination of sugars and related substances.Anal Chem,1956,28:350-356.
15 Kawauchi S,Iwanaga S,Samejima Y,et al.Isolation and characterization of two phospholipase A's from the venom of Agkistrodon lays blomhoffii.Biochim Biophys Acta,1971,236:142-160.
16 Satake M,Murata Y,Suzuki T.Studies on snake venom.XIII.Chromatographic separation and properties of three proteinases from Agkistrodon halys blomhoffii venom.J Biochem (Tokyo),1963,53:438-447.
17 Kondo H,Kondo S,Ikezawa H,et al.Studies on the quantitative method for determination of hemorrhagic activity of Habu snake venom.Jpn J Med Sci Biol,1960,13:43-52.

[1]	杨勇勋, 祝雅庭, 罗伦才, 耿浩. 岩须精油的化学成分及抗凝血活性研究[J]. 天然产物研究与开发, 2024, 36(增刊1): 59-63.
[2]	杨勇勋, 颜瑜, 黄海燕. 红棕杜鹃（原变种）叶中一个新的杜鹃色烯杂萜及其抗凝血活性评价[J]. 天然产物研究与开发, 2023, 35(4): 607-611.
[3]	闫兆威, 张敏, 魏明刚, 姚鑫, 刘江云. 天然木脂素苷类成分eleutheroside E对膝骨性关节炎的治疗作用及其与MMPs相互作用方式的分析[J]. 天然产物研究与开发, 2021, 33(8): 1339-1347.
[4]	熊雅茹, 傅红, 杨方, . 阿胶多肽的高分辨质谱鉴定及活性研究[J]. 天然产物研究与开发, 2020, 32(8): 1348-1356.
[5]	王风楼1，邓文娟2，贺改英1*. 黄芪复方对成纤维细胞胶原蛋白和MMP的影响[J]. 天然产物研究与开发, 2019, 31(增刊2): 84-87.
[6]	郭静1,2，李敏3，杨毅2，张霖2，孙黔云1,2. 绿原酸对小鼠急性肺损伤的保护作用研究[J]. 天然产物研究与开发, 2018, 30(7): 1214-1218.
[7]	王海丽，陈毅，肖林焱，薛露，单鸣秋，张丽，丁安伟*. 茜草炭对急性血瘀模型大鼠抗凝血倾向的活性部位筛选[J]. 天然产物研究与开发, 2016, 28(11): 1797-1801.
[8]	夏梦媛1,2，王俐1，王跃虎2. 胡椒属植物抗血小板和抗血栓活性研究进展[J]. 天然产物研究与开发, 2016, 28(10): 1676-1685.
[9]	文彬，陈然，彭佩纯，邓鑫*. 天然牛磺酸对肝硬化大鼠MMP-9、Integrin-β1表达的调控[J]. 天然产物研究与开发, 2014, 26(12): 1939-1943.

蕲蛇蛇毒中一种新型金属蛋白酶AA-MP-I的纯化和表征

Purification and Characterization of a Novel Metalloproteinase AA-MP-I from the Venom of Agkistrodon acutus

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 9

编辑推荐

Metrics

本文评价