Send to

Choose Destination
Chem Biol Drug Des. 2018 Nov 27. doi: 10.1111/cbdd.13450. [Epub ahead of print]

Characterization and regulation of MT1-MMP cell surface-associated activity.

Author information

Departments of Chemistry and Biology, Torrey Pines Institute for Molecular Studies, Port St. Lucie, FL, 34987, USA.
Department of Chemistry & Biochemistry, Florida Atlantic University, Jupiter, FL, 33458, USA.
Departments of Medicine/Cancer Prevention and Pathology, Stony Brook University, Stony Brook, NY, 11794, USA.
Cancer Research Center, Sanford Burnham Prebys Medical Research Institute, La Jolla, CA, 92037, USA.
Department of Pathology and the Karmanos Cancer Institute, Wayne State University, Detroit, MI, 48201, USA.
Division of Molecular Medicine & Genetics, Department of Internal Medicine, and the Life Sciences Institute, University of Michigan, Ann Arbor, MI, 48109, USA.
The Scripps Research Institute/Scripps Florida, Jupiter, FL, 33458, USA.


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 towards 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. This article is protected by copyright. All rights reserved.


Cell Surface Proteolysis; Cell-based Assay; Collagenolysis; Matrix Metalloproteinase; Protease Inhibitor


Supplemental Content

Full text links

Icon for Wiley
Loading ...
Support Center