期刊文献+
共找到1篇文章
< 1 >
每页显示 20 50 100
Gelatin degradation assay reveals MMP-9 inhibitors and function of O-glycosylated domain 被引量:3
1
作者 Jennifer Vandooren Nathalie Geurts +4 位作者 Erik Martens Philippe E Van den Steen Steven De Jonghe piet herdewijn Ghislain Opdenakker 《World Journal of Biological Chemistry》 CAS 2011年第1期14-24,共11页
AIM: To establish a novel, sensitive and high-throughput gelatinolytic assay to define new inhibitors and compare domain deletion mutants of gelatinase B/matrix metalloproteinase (MMP)-9. METHODS: Fluorogenic Dye-quen... AIM: To establish a novel, sensitive and high-throughput gelatinolytic assay to define new inhibitors and compare domain deletion mutants of gelatinase B/matrix metalloproteinase (MMP)-9. METHODS: Fluorogenic Dye-quenched (DQ)TM-gelatin was used as a substrate and biochemical parameters (substrate and enzyme concentrations, DMSO solvent concentrations) were optimized to establish a highthroughput assay system. Various small-sized libraries (ChemDiv, InterBioScreen and ChemBridge) of hetero-cyclic, drug-like substances were tested and compared with prototypic inhibitors. RESULTS: First, we designed a test system with gelatin as a natural substrate. Second, the assay was validated by selecting a novel pyrimidine-2,4,6-trione (barbitu- rate) inhibitor. Third, and in line with present structural data on collagenolysis, it was found that deletion of the O-glycosylated region significantly decreased gelatinolytic activity (kcat/kM ± 40% less than full-length MMP-9). CONCLUSION: The DQTM-gelatin assay is useful in high-throughput drug screening and exosite targeting. We demonstrate that flexibility between the catalytic and hemopexin domain is functionally critical for gelatinolysis. 展开更多
关键词 Exosite INHIBITORS FLUOROGENIC SUBSTRATE GELATIN High-throughput screening assays Matrix metalloproteinase-9 SUBSTRATE specificity
下载PDF
上一页 1 下一页 到第
使用帮助 返回顶部