来自竹叶青蛇毒液的一种类凝血酶的特性研究。

Characterization of a thrombin-like enzyme from the venom of Trimeresurus jerdonii.

作者信息

Lu Q M, Jin Y, Li D S, Wang W Y, Xiong Y L

机构信息

Department of Animal Toxinology, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, Yunnan, China.

出版信息

Toxicon. 2000 Sep;38(9):1225-36. doi: 10.1016/s0041-0101(99)00222-6.

DOI:10.1016/s0041-0101(99)00222-6

PMID:10736476

Abstract

From the venom of Trimeresurus jerdonii, a distinct thrombin-like enzyme, called jerdonobin, was purified by DEAE A-25 ion-exchange chromatography, Sephadex G-75 gel filtration, and fast protein liquid chromatography (FPLC). SDS-PAGE analysis of this enzyme shows that it consists of a single polypeptide chain with a molecular weight of 38,000. The NH(2)-terminal amino acid sequence of jerdonobin has great homology with venom thrombin-like enzymes documented. Jerdonobin is able to hydrolyze several chromogenic substrates. The enzyme directly clots fibrinogen with an activity of 217 NIH units/mg. The fibrinopeptides released, identified by HPLC, consisted of fibrinopeptide A and a small amount of fibrinopepide B. The activities of the enzyme were inhibited by phenylmethylsulfonyl fluoride (PMSF) and p-nitrophenyl-p-guanidinobenzoate (NPGB). However, metal chelator (EDTA) had no effect on it, indicating it is venom serine protease.

摘要

从竹叶青蛇（Trimeresurus jerdonii）的毒液中，通过DEAE A - 25离子交换色谱法、Sephadex G - 75凝胶过滤法和快速蛋白质液相色谱法（FPLC）纯化出一种独特的类凝血酶酶，称为吉氏凝血酶（jerdonobin）。对该酶的SDS - PAGE分析表明，它由一条分子量为38,000的单多肽链组成。吉氏凝血酶的NH(2)-末端氨基酸序列与已记录的毒液类凝血酶具有高度同源性。吉氏凝血酶能够水解几种显色底物。该酶能直接使纤维蛋白原凝固，活性为217 NIH单位/毫克。通过HPLC鉴定释放的纤维蛋白肽，其由纤维蛋白肽A和少量纤维蛋白肽B组成。该酶的活性受到苯甲基磺酰氟（PMSF）和对硝基苯基 - p - 胍基苯甲酸酯（NPGB）的抑制。然而，金属螯合剂（EDTA）对其没有影响，表明它是一种毒液丝氨酸蛋白酶。