Intermolecular Autolytic Cleavage Can Contribute to the Activation of Progelatinase A by Cell Membranes (*)
- Susan J. Atkinson(1)(§),
- Thomas Crabbe(2),
- Susan Cowell(1),
- Robin V. Ward(1)(¶),
- Michael J. Butler(1),
- Hiroshi Sato(3),
- Motoharu Seiki(3),
- John J. Reynolds(1) and
- Gillian Murphy(1)
- From the (1)Department of Cell and Molecular Biology, Strangeways Research Laboratory, Cambridge CB1 4RN, United Kingdom,
- (2)Celltech Research, Slough, Berks SL1 4EN, United Kingdom, and the
- (3)Department of Molecular Virology and Oncology, University of Kanazawa, Cancer Research Institute, 13-1 Takara-machi Kanazawa, Ishikawa 920, Japan
- § To whom correspondence should be addressed.
Abstract
Membrane-type matrix metalloproteinase (MT-MMP) messenger RNA and protein expression were shown to be elevated in human fibroblasts following treatment with concanavalin A, coincident with the induction of the ability to process progelatinase A.
CHO cells transfected with the cDNA for MT-MMP were able to process both wild type progelatinase A and a catalytically inactive mutant, E375A progelatinase A. Both proenzymes were converted to a 68-kDa intermediate (reducing gels) form, but only the wild type enzyme was processed further to a 66-kDa end product. In contrast, both forms of progelatinase were processed via the 68-kDa intermediate to 66 kDa by concanavalin A-stimulated fibroblasts.
Further study of the processing of E375A progelatinase A by plasma membrane preparations from concanavalin A-stimulated fibroblasts showed that addition of active gelatinase A enhanced processing to the mature form. It was concluded that cell membrane-mediated activation of progelatinase A could be via a cascade involving both MT-MMP and intermolecular autolytic cleavage.
Footnotes
-
↵* This work was supported by the Medical Research Council and the Arthritis and Rheumatism Council. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
-
↵1 The abbreviations used are:
- MT-MMP
-
membrane-type matrix metalloproteinase
- CHO
-
Chinese hamster ovary fibroblasts
- PAGE
-
polyacrylamide gel electrophoresis
- APMA
-
p-aminophenylmercuric acetate
- DMEM
-
Dulbecco's modified Eagle's medium
- TIMP1 and −2
-
tissue inhibitors of metalloproteinases
- ECL
-
enhanced chemiluminescence
- ConA
-
concanavalin A.
-
↵2H. Will and G. Murphy, unpublished data.
-
↵3S. Atkinson and R. Hembry, unpublished data.
-
- Received May 31, 1995.
- Revision received September 28, 1995.
- © 1995 by The American Society for Biochemistry and Molecular Biology, Inc.
