Structural and mechanistic insights into collagen degradation by a bacterial collagenolytic serine protease in the subtilisin family
Structural and mechanistic insights into collagen degradation by a bacterial collagenolytic serine protease in the subtilisin family
复制标题
枯草杆菌蛋白酶家族中细菌溶胶丝氨酸蛋白酶对胶原蛋白降解的结构和机制见解
DOI：
10.1111/mmi.12412
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发表时间：
2013-12-01
影响因子：
3.6
通讯作者：
Zhang, Yu-Zhong
中科院分区：
文献类型：
--
作者：
Ran, Li-Yuan;Su, Hai-Nan;Zhang, Yu-Zhong
已报道来源于环境或病原微生物的枯草杆菌蛋白酶家族中的许多蛋白酶是胶原分解丝氨酸蛋白酶。然而，其胶原降解机制仍不清楚。在此，研究了来自交替假单胞菌属SM 9913的S8胶原蛋白溶解蛋白酶MCP-01对I型胶原纤维的降解机制。原子力显微镜观察和生化分析证实，MCP-01主要通过水解胶原纤维中的蛋白聚糖和端肽，从胶原纤维中逐渐释放单原纤维并从原纤维中释放胶原单体。结构和突变分析表明，主要由环7、9和11组成的扩大的底物结合口袋是胶原蛋白识别所必需的，并且这些环上的酸性和芳香族残基形成了带负电荷的疏水环境，用于胶原蛋白结合。MCP-01对胶原蛋白中P1位点的Pro或碱性残基和/或P1'位点的Gly的肽键显示出非严格的偏好。His 211是MCP-01的P1-碱性-残基偏好的关键残基。我们的研究提供了结构和机制的见解，胶原蛋白降解的S8胶原蛋白水解酶，这是有助于开发治疗疾病的S8胶原蛋白水解酶作为致病因素，并在研究环境有机氮降解机制。
A number of proteases in the subtilisin family derived from environmental or pathogenic microorganisms have been reported to be collagenolytic serine proteases. However, their collagen degradation mechanisms remain unclear. Here, the degradation mechanism of type I collagen fibres by the S8 collagenolytic protease MCP‐01, from Pseudoalteromonas sp. SM9913, was studied. Atomic force microscopy observation and biochemical analysis confirmed that MCP‐01 progressively released single fibrils from collagen fibres and released collagen monomers from fibrils mainly by hydrolysing proteoglycans and telopeptides in the collagen fibres. Structural and mutational analyses indicated that an enlarged substrate‐binding pocket, mainly composed of loops 7, 9 and 11, is necessary for collagen recognition and that the acidic and aromatic residues on these loops form a negatively charged, hydrophobic environment for collagen binding. MCP‐01 displayed a non‐strict preference for peptide bonds with Pro or basic residues at the P1 site and/or Gly at the P1’ site in collagen. His211 is a key residue for the P1‐basic‐residue preference of MCP‐01. Our study gives structural and mechanistic insights into collagen degradation of the S8 collagenolytic protease, which is helpful in developing therapeutics for diseases with S8 collagenolytic proteases as pathogenic factors and in studying environmental organic nitrogen degradation mechanisms.