Identification of the active site of gelatinase B as the structural element sufficient for converting a protein to a metalloprotease.
Identification of the active site of gelatinase B as the structural element sufficient for converting a protein to a metalloprotease.
复制标题
鉴定明胶酶 B 的活性位点作为足以将蛋白质转化为金属蛋白酶的结构元件。
DOI：
10.1021/bi015930p
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发表时间：
2002
期刊：
影响因子：
2.9
通讯作者：
Pourmotabbed,Tayebeh
中科院分区：
文献类型：
--
作者：
Kaur,Kuljeet;Zhu,Kijeen;Whittemore,MarilynS;Petersen,RichardL;Lichte,Andrea;Tschesche,Harald;Pourmotabbed,Tayebeh
明胶酶B是基质金属蛋白酶家族的成员，其有效地切割明胶、弹性蛋白以及V型和X型胶原。为了理解酶的活性位点（氨基酸残基373 - 456）在这些活性中的贡献，我们研究了由麦芽糖结合蛋白和明胶酶B的活性位点区域组成的融合蛋白的催化性质。我们发现，在麦芽糖结合蛋白中加入明胶酶B的活性位点（相当于总蛋白分子的12%），足以赋予该蛋白切割肽底物Mca-PLGL（Dpa）AR-NH 2和DNP-PLGLWA-（d）-R-NH 2的能力。该融合蛋白水解Mca-PLGL（Dpa）AR-NH 2肽的效率与基质溶解素相当，kcat/Km为1.07 × 106 M-1h-1。然而，融合蛋白不能降解大底物明胶。EDTA对该蛋白活性的抑制表明其活性具有金属依赖性。ESR分析表明，融合蛋白与一分子Zn ~（2+）结合。此外，Z-Pro-Leu-Gly-hydroxamate和TIMP-1抑制该蛋白的活性，表明该融合蛋白的活性位点的结构类似于其他金属蛋白酶。这些数据提供了关于将蛋白质转化为金属蛋白酶所需的结构元件的基本信息。
Gelatinase B is a member of the matrix metalloproteinase family that efficiently cleaves gelatin, elastin, and types V and X collagen. To understand the contribution of the active site of the enzyme (amino acid residues 373−456) in these activities, we studied catalytic properties of a fusion protein consisting of maltose binding protein and the active site region of gelatinase B. We found that addition of the active site of gelatinase B, which corresponds to 12% of the total protein molecule, to maltose binding protein is sufficient to endow the protein with the ability to cleave the peptide substrates Mca-PLGL(Dpa)AR-NH2and DNP-PLGLWA-(d)-R-NH2. The fusion protein hydrolyzed the Mca-PLGL(Dpa)AR-NH2peptide with the same efficiency as that of the stromelysin,kcat/Km≃ 1.07 × 106M-1h-1. The fusion protein, however, was not able to degrade the large substrate, gelatin. Inhibition of the activity of the protein by EDTA suggested that its activity was metal dependent. ESR analyses indicated that the fusion protein bound one molecule of Zn2+. In addition, Z-Pro-Leu-Gly-hydroxamate and TIMP-1 inhibited the activity of the protein, suggesting that the structure of the active site of the fusion protein is similar to that of the other metalloproteinases. These data provide fundamental information about the structural elements required for transforming a protein to a metalloprotease.
DOI：
--
发表时间：
1991
期刊：
影响因子：
--
作者：
M. Mareel;P. Baetselier;F. Roy
通讯作者：
F. Roy
影响因子：
4.1
作者：
CAWSTON, TE;MURPHY, G;REYNOLDS, JJ
通讯作者：
REYNOLDS, JJ