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- W2902339978 endingPage "1264" @default.
- W2902339978 startingPage "1251" @default.
- W2902339978 abstract "Quantitative assessment of MT1-MMP cell surface-associated proteolytic activity remains undefined. Presently, MT1-MMP was stably expressed and a cell-based FRET assay developed to quantify activity toward synthetic collagen-model triple-helices. To estimate the importance of cell surface localization and specific structural domains on MT1-MMP proteolysis, activity measurements were performed using a series of membrane-anchored MT1-MMP mutants and compared directly with those of soluble MT1-MMP. MT1-MMP activity (kcat /KM ) on the cell surface was 4.8-fold lower compared with soluble MT1-MMP, with the effect largely manifested in kcat . Deletion of the MT1-MMP cytoplasmic tail enhanced cell surface activity, with both kcat and KM values affected, while deletion of the hemopexin-like domain negatively impacted KM and increased kcat . Overall, cell surface localization of MT1-MMP restricts substrate binding and protein-coupled motions (based on changes in both kcat and KM ) for catalysis. Comparison of soluble and cell surface-bound MT2-MMP revealed 12.9-fold lower activity on the cell surface. The cell-based assay was utilized for small molecule and triple-helical transition state analog MMP inhibitors, which were found to function similarly in solution and at the cell surface. These studies provide the first quantitative assessments of MT1-MMP activity and inhibition in the native cellular environment of the enzyme." @default.
- W2902339978 created "2018-12-11" @default.
- W2902339978 creator A5021912229 @default.
- W2902339978 creator A5023401314 @default.
- W2902339978 creator A5030012912 @default.
- W2902339978 creator A5048088279 @default.
- W2902339978 creator A5063462261 @default.
- W2902339978 creator A5068707393 @default.
- W2902339978 creator A5077751051 @default.
- W2902339978 creator A5080476399 @default.
- W2902339978 date "2018-12-19" @default.
- W2902339978 modified "2023-10-14" @default.
- W2902339978 title "Characterization and regulation of <scp>MT</scp>1‐<scp>MMP</scp> cell surface‐associated activity" @default.
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