Complex pattern of membrane type 1 matrix metalloproteinase shedding. Regulation by autocatalytic cell surface inactivation of active enzyme

Marta Toth, Sonia Hernandez-Barrantes, Pamela Osenkowski, M. Margarida Bernardo, David C. Gervasi, Yoichiro Shimura, Oussama Meroueh, Lakshmi P. Kotra, Beatriz G. Gálvez, Alicia G. Arroyo, Shahriar Mobashery, Rafael Fridman

Research output: Contribution to journalArticle

99 Scopus citations

Abstract

Membrane type 1 matrix metalloproteinase (MT1-MMP) is a type I transmembrane MMP shown to play a critical role in normal development and in malignant processes. Emerging evidence indicates that MT1-MMP is regulated by a process of ectodomain shedding. Active MT1-MMP undergoes autocatalytic processing on the cell surface, leading to the formation of an inactive 44-kDa fragment and release of the entire catalytic domain. Analysis of the released MT1-MMP forms in various cell types revealed a complex pattern of shedding involving two major fragments of 50 and 18 kDa and two minor species of 56 and 31-35 kDa. Protease inhibitor studies and a catalytically inactive MT1-MMP mutant revealed both autocatalytic (18 kDa) and non-autocatalytic (56, 50, and 31-35 kDa) shedding mechanisms. Purification and sequencing of the 18-kDa fragment indicated that it extends from Tyr112 to Ala255. Structural and sequencing data indicate that shedding of the 18-kDa fragment is initiated at the Gly284-Gly285 site, followed by cleavage between the conserved Ala255 and Ile256 residues near the conserved methionine turn, a structural feature of the catalytic domain of all MMPs. Consistently, a recombinant 18-kDa fragment had no catalytic activity and did not bind TIMP-2. Thus, autocatalytic shedding evolved as a specific mechanism to terminate MT1-MMP activity on the cell surface by disrupting enzyme integrity at a vital structural site. In contrast, functional data suggest that the non-autocatalytic shedding generates soluble active MT1-MMP species capable of binding TIMP-2. These studies suggest that ectodomain shedding regulates the pericellular and extracellular activities of MT1-MMP through a delicate balance of active and inactive enzyme-soluble fragments.

Original languageEnglish (US)
Pages (from-to)26340-26350
Number of pages11
JournalJournal of Biological Chemistry
Volume277
Issue number29
DOIs
StatePublished - Jul 19 2002
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Toth, M., Hernandez-Barrantes, S., Osenkowski, P., Margarida Bernardo, M., Gervasi, D. C., Shimura, Y., Meroueh, O., Kotra, L. P., Gálvez, B. G., Arroyo, A. G., Mobashery, S., & Fridman, R. (2002). Complex pattern of membrane type 1 matrix metalloproteinase shedding. Regulation by autocatalytic cell surface inactivation of active enzyme. Journal of Biological Chemistry, 277(29), 26340-26350. https://doi.org/10.1074/jbc.M200655200