斯氏植滴虫中的细胞外金属蛋白酶

Extracellular metalloproteinases in Phytomonas serpens.

作者信息

Vermelho Alane B, Almeida Flávia V S, Bronzato Leandro S, Branquinha Marta H

机构信息

Departamento de Microbiologia Geral, Instituto Microbiologia Prof. Paulo de Góes, Centro de Ciências de Saúde, Bl. I, Cidade Universitária, Universidade Federal do Rio de Janeiro, 21941-590, Rio de Janeiro, Brazil.

出版信息

Can J Microbiol. 2003 Mar;49(3):221-4. doi: 10.1139/w03-020.

DOI:10.1139/w03-020

PMID:12795409

Abstract

The detection of extracellular proteinases in Phytomonas serpens, a trypanosomatid isolated from tomato fruits, is demonstrated in this paper. Maximal production occurred at the end of the logarithmic phase of growth. These enzymes exhibited selective substrate utilization in SDS-PAGE, being more active with gelatin; hemoglobin and bovine serum albumin were not degraded. Three proteinases were detected in SDS-PAGE-gelatin, with apparent molecular masses between 94 and 70 kDa. The proteolytic activity was completely blocked by 1,10-phenanthroline and strongly inhibited by EDTA, whereas a partial inhibition was observed with trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64) and soybean trypsin inhibitor; phenylmethylsulfonyl fluoride weakly inhibited the enzymes. This inhibition profile indicated that these extracellular proteinases belong to the metalloproteinase class.

摘要

本文展示了从番茄果实中分离出的锥虫——斯氏植鞭毛虫（Phytomonas serpens）细胞外蛋白酶的检测情况。最大产量出现在生长对数期结束时。这些酶在SDS-PAGE中表现出选择性底物利用，对明胶更具活性；血红蛋白和牛血清白蛋白未被降解。在SDS-PAGE-明胶中检测到三种蛋白酶，其表观分子量在94至70 kDa之间。蛋白水解活性被1,10-菲咯啉完全阻断，并被EDTA强烈抑制，而反式环氧琥珀酰-L-亮氨酰胺基-（4-胍基）丁烷（E-64）和大豆胰蛋白酶抑制剂则有部分抑制作用；苯甲基磺酰氟对这些酶有微弱抑制作用。这种抑制模式表明这些细胞外蛋白酶属于金属蛋白酶类。