Molecular cloning and biochemical characterisation of proteases from Staphylococcus epidermidis
Molecular cloning and biochemical characterisation of proteases from Staphylococcus epidermidis
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DOI：
10.1515/bc.2001.192
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发表时间：
2001-11-01
影响因子：
3.7
通讯作者：
Dubin, A
中科院分区：
文献类型：
--
作者：
Dubin, G;Chmiel, D;Dubin, A
本文报道了表皮葡萄球菌胞外半胱氨酸（Ecp）和丝氨酸（Esp）蛋白酶的完整编码序列和部分氨基酸序列（经化学测序确定）。第一种酶显示出与金黄色葡萄球菌半胱氨酸蛋白酶（葡萄球菌蛋白酶）的延伸的序列相似性，第二种酶类似于由该物种产生的丝氨酸蛋白酶。在两种酶中编码成熟蛋白的序列的直接上游区域分别显示出与由sspB和sspA编码的前片段的显著同源性，从而表明所表征的酶也可以作为前蛋白产生。此外，我们报告的半胱氨酸蛋白酶的一些生物学特性，有助于更好地了解其作为一个可能的毒力因子的作用。该酶的蛋白水解活性可被人α-2-巨球蛋白快速有效地抑制;然而，人激肽原以及半胱氨酸蛋白酶抑制剂（A、C和D）不具有抑制作用。此外，该蛋白酶能够通过有限的蛋白水解使α-1-抗胰蛋白酶和HMW-激肽原失活，但既不能使α-1-抗胰凝乳蛋白酶失活，也不能使抗凝血酶III失活。
We report the complete coding sequence and the partial amino acid sequence (determined by chemical sequencing) of Staphylococcus epidermidis extracellular cysteine (Ecp) and serine (Esp) proteases. The first enzyme shows an extended sequence similarity to Staphylococcus aureus cysteine protease (staphopain) and the second one resembles the serine protease produced by that species. The region directly upstream of the sequence coding for the mature protein in both enzymes displays significant homology to the profragments encoded by sspB and sspA, respectively, thus suggesting that the characterised enzymes may also be produced as proproteins. Furthermore, we report some biological properties of the cysteine protease, contributing to a better understanding of its role as a possible virulence factor. The proteolytic activity of this enzyme was rapidly and efficiently inhibited by human alpha -2-macroglobulin; however, human kininogen as well as cystatins (A, C and D) were not inhibitory. Moreover, the protease was capable of inactivating, by limited proteolysis, both a-l-antitrypsin and HMW-kininogen, but neither alpha -1-antichymotrypsin nor antithrombin Ill.