Regulation of membrane-type matrix metalloproteinase 1 activity by dynamin-mediated endocytosis
- *Department of Pharmacology, University of Minnesota, 6-120 Jackson Hall, 321 Church Street SE, Minneapolis, MN 55455; †Departments of Pathology and Virology, Haartman Institute, University of Helsinki and University Hospital, FIN-00014 Helsinki, Finland; and ‡Comprehensive Cancer Center and Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109
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Edited by Marilyn Gist Farquhar, University of California at San Diego, La Jolla, CA, and approved September 20, 2001 (received for review June 12, 2001)
Abstract
Membrane-type matrix metalloproteinase 1 (MT1-MMP) plays a critical role in extracellular matrix remodeling under both physiological and pathological conditions. However, the mechanisms controlling its activity on the cell surface remain poorly understood. In this study, we demonstrate that MT1-MMP is regulated by endocytosis. First, we determined that Con A induces proMMP-2 activation in HT1080 cells by shifting endogenous MT1-MMP from intracellular compartments to cell surface. This phenotype was mimicked by the cytoplasmic truncation mutant MT1ΔC with more robust pro-MMP-2 activation and cell surface expression than wild-type MT1-MMP in transfected cells. MT1ΔC was subsequently shown to be resistant to Con A treatment whereas MT1-MMP remains competent, suggesting that Con A regulates MT1-MMP activity through cytoplasmic domain-dependent trafficking. Indeed, MT1-MMP was colocalized with clathrin on the plasma membrane and with endosomal antigen 1 in endosomes. Internalization experiments revealed that MT1-MMP is internalized rapidly in clathrin-coated vesicles whereas MT1ΔC remains on cell surface. Coexpression of a dominant negative mutant of dynamin, K44A, resulted in elevation of MT1-MMP activity by interfering with the endocytic process. Thus, MT1-MMP is regulated by dynamin-dependent endocytosis in clathrin-coated pits through its cytoplasmic domain.
Footnotes
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↵ § To whom reprint requests should be addressed. E-mail: peixx003{at}tc.umn.edu.
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This paper was submitted directly (Track II) to the PNAS office.
- Abbreviations:
- MT-MMP,
- membrane-type matrix metalloproteinase;
- MMP,
- matrix metalloproteinase;
- ECM,
- extracellular matrix;
- EEA1,
- early endosomal antigen 1;
- wt,
- wild type;
- MDCK,
- Madin–Darby canine kidney
- Copyright © 2001, The National Academy of Sciences
