Substrate hydrolysis by matrix metalloproteinase-9
Substrate hydrolysis by matrix metalloproteinase-9
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DOI：
10.1074/jbc.m100900200
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发表时间：
2001-06-08
影响因子：
4.8
通讯作者：
Smith, JM
中科院分区：
文献类型：
--
作者：
Kridel, SJ;Chen, E;Smith, JM
所有基质金属蛋白酶（MMPs）的催化裂缝具有相似的结构，这引发了关于蛋白质家族中底物识别冗余的问题。在本研究中，应用无偏噬菌体展示策略来定义MMP-9的底物识别谱。确定了三组底物，每组占据催化口袋内的一组不同的子位点。最普遍的基序在P-3至P-2 '处含有序列Pro-X-X-Hy-（Ser/Thr）。该序列与胶原内的MMP切割位点相似，并且与已经为其他MMP选择的底物同源。尽管有这种相似性，但这里鉴定的大多数底物对MMP-9的选择性超过MMP-7和MMP-13。这一观察结果表明，底物选择性是由P-3和P-1 '以外位置的关键亚位点相互作用赋予的。该研究表明MMP-9在P-2和P-1都具有对Arg的独特偏好，并且在P-2 '具有对Ser/Thr的偏好。使用含有共有MMP-9识别基序的底物来查询蛋白质数据库。一个令人惊讶的有限的名单推定的生理基板被确定。这些蛋白质的功能影响导致关于MMP-9的生理底物的可检验的假设。
The catalytic clefts of all matrix metalloproteinases (MMPs) have a similar architecture, raising questions about the redundancy in substrate recognition across the protein family. In the present study, an unbiased phage display strategy was applied to define the substrate recognition profile of MMP-9. Three groups of substrates were identified, each occupying a distinct set of subsites within the catalytic pocket. The most prevalent motif contains the sequence Pro-X-X-Hy-(Ser/Thr) at P-3 through P-2'. This sequence is similar to the MMP cleavage sites within the collagens and is homologous to substrates the have been selected for other MMPs. Despite this similarity, most of the substrates identified here are selective for MMP-9 over MMP-7 and MMP-13. This observation indicates that substrate selectivity is conferred by key subsite interactions at positions other than P-3 and P-1'. This study shows that MMP-9 has a unique preference for Arg at both P-2 and P-1, and a preference for Ser/Thr at P-2'. Substrates containing the consensus MMP-9 recognition motif were used to query the protein data bases. A surprisingly limited List of putative physiologic substrates was identified. The functional implications of these proteins lead to testable hypotheses regarding physiologic substrates for MMP-9.