Characterization and molecular cloning of a glutamyl endopeptidase from Staphylococcus epidermidis
Characterization and molecular cloning of a glutamyl endopeptidase from Staphylococcus epidermidis
复制标题
DOI：
10.1006/mpat.2002.0515
复制
发表时间：
2002-07-01
影响因子：
3.8
通讯作者：
Kimura, S
中科院分区：
文献类型：
--
作者：
Ohara-Nemoto, Y;Ikeda, Y;Kimura, S
采用透析膜技术从表皮葡萄球菌ATCC 14990中纯化出一种新型胞外肽链内切酶，命名为GluSE，并克隆了其结构基因（gseA）。 GluSE 是一种 27 kDa 的谷氨酸特异性蛋白酶，最适 pH 为 8.0。蛋白水解活性被氟磷酸二异丙酯特异性抑制，表明它是一种丝氨酸蛋白酶。 gseA编码282个氨基酸的单一多肽，推导分子量为30,809，其中前19个N端氨基酸与从Val-67开始的推导序列完全匹配，表明GluSE是用前肽合成的。 GluSE 的氨基酸序列与金黄色葡萄球菌 V8 蛋白酶 (GluV8) 具有 50.5% 的同一性。尽管 GluSE 缺少 GluV8 的 PBN/PBZ 三肽的 C 端 12 个重复，但 His-117、Asp-159 和 Ser-235 的催化三联体在 GluSE 中是保守的。 Southern 杂交分析表明，gseA 在染色体 DNA 上以单个拷贝的形式存在。在透析膜技术的贴壁培养条件下明显观察到 GluSE 的产生，但在浮游培养条件下没有观察到 GluSE 的产生，这一发现强烈表明 GluSE 可能参与表皮葡萄球菌感染导致多种组织损伤的重要病因过程。 (C) 2002 Elsevier Science Ltd. 保留所有权利。
A novel extracellular endopeptidase, designated GluSE, was purified from Staphylococcus epidermidis ATCC 14990 cultured by the dialysis membrane technique, and the structural gene (gseA) was cloned. GluSE was a 27 kDa, glutamic acid-specific protease, and the optimal pH was 8.0. The proteolytic activity was specifically inhibited with diisopropyl fluorophosphate, indicating that it is a serine protease. The gseA encoded a single polypeptide of 282 amino acids with a deduced molecular weight of 30,809, in which the first 19 N-terminal amino acids completely matched the deduced sequence starting at Val-67, suggesting that GluSE is synthesized with a propeptide. The amino acid sequence of GluSE exhibited 50.5% identity to Staphylococcus aureus V8-protease (GluV8). Although GluSE lacks a C-terminal 12 repeats of the PBN/PBZ tripeptide of GluV8, a catalytic triad of His-117, Asp-159 and Ser-235 was conserved in GluSE. Southern hybridization analysis revealed that gseA exists as a single copy on the chromosomal DNA. The finding that production of GluSE was obviously observed in the adherent culture conditions of the dialysis membrane technique, but not in the planktonic culture conditions, strongly suggests that GluSE could be involved in an important etiologic process in S. epidermidis infection leading to multiple tissue damages. (C) 2002 Elsevier Science Ltd. All rights reserved.