Functional Interplay between Type I Collagen and Cell Surface Matrix Metalloproteinase Activity

doi:10.1074/jbc.M005631200

Functional Interplay between Type I Collagen and Cell Surface Matrix Metalloproteinase Activity*

From the ^‡Departments of Cell and Molecular Biology and Obstetrics and Gynecology, Northwestern University Medical School, Chicago, Illinois 60611 and the ^§Faculty of Dentistry and Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada

Abstract

Type I collagen stimulation of pro-matrix metalloproteinase (pro-MMP)-2 activation by ovarian cancer cells involves β₁ integrin receptor clustering; however, the specific cellular and biochemical events that accompany MMP processing are not well characterized. Collagenolysis is not required for stimulation of pro-MMP-2 activation, and denatured collagen does not elicit an MMP-2 activation response. Similarly, DOV13 cells bind to intact collagen utilizing both α₂β₁ and α₃β₁ integrins but interact poorly with collagenase-treated or thermally denatured collagen. Antibody-induced clustering of α₃β₁ strongly promotes activation of pro-MMP-2, whereas α₂β₁integrin clustering has only marginal effects. Membrane-type 1 (MT1)-MMP is present on the DOV13 cell surface as both an active 55-kDa TIMP-2-binding species and a stable catalytically inactive 43-kDa form. Integrin clustering stimulates cell surface expression of MT1-MMP and co-localization of the proteinase to aggregated integrin complexes. Furthermore, cell surface proteolysis of the 55-kDa MT1-MMP species occurs in the absence of active MMP-2, suggesting MT1-MMP autolysis. Cellular invasion of type I collagen matrices requires collagenase activity, is blocked by tissue inhibitor of metalloproteinases-2 (TIMP-2) and collagenase-resistant collagen, is unaffected by TIMP-1, and is accompanied by pro-MMP-2 activation. Together, these data indicate that integrin stimulation of MT1-MMP activity is a rate-limiting step for type I collagen invasion and provide a mechanism by which this activity can be down-regulated following collagen clearance.

Footnotes

↵* This work was supported by National Institutes of Health Training Grant 5T32 GM08061 (to S. M. E.), United States Army MRMC Training Grant DAMD170010386 (to Y. I. W.), and NCI Research Grant RO1 CA86984 (to M. S. S.) from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¶ To whom correspondence should be addressed: Dept. of Cell and Molecular Biology, Northwestern University Medical School, 303 E. Chicago Ave., Tarry 8-715, Chicago, IL 60611. Tel.: 312-908-8216; Fax: 312-503-7912; E-mail: mss130@northwestern.edu.
Published, JBC Papers in Press, April 30, 2001, DOI 10.1074/jbc.M005631200
Abbreviations:

MMP

matrix metalloproteinase

MT1-MMP

membrane type-1 matrix metalloproteinase

TIMP

tissue inhibitor of matrix metalloproteinase

ConA

concanavalin A

rCBD123

recombinant collagen binding domain

rCD

recombinant carboxyl hemopexin domain

PBS

phosphate-buffered saline

TBS

Tris-buffered saline

BSA

bovine serum albumin

mAb

monoclonal antibody

CR

collagenase-resistant

MES

4-morpholineethanesulfonic acid

MMPI

MMP inhibitor

CHO

Chinese hamster ovary
- Received June 27, 2000.
- Revision received April 4, 2001.
The American Society for Biochemistry and Molecular Biology, Inc.