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

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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

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Matrix Metalloproteinase 14
Enzymes
Tissue Inhibitor of Metalloproteinase-2
Matrix Metalloproteinases
Catalytic Domain
Protease Inhibitors
Methionine
Purification
Catalyst activity

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Toth, M., Hernandez-Barrantes, S., Osenkowski, P., Margarida Bernardo, M., Gervasi, D. C., Shimura, Y., ... 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

Complex pattern of membrane type 1 matrix metalloproteinase shedding. Regulation by autocatalytic cell surface inactivation of active enzyme. / Toth, Marta; Hernandez-Barrantes, Sonia; Osenkowski, Pamela; Margarida Bernardo, M.; Gervasi, David C.; Shimura, Yoichiro; Meroueh, Oussama; Kotra, Lakshmi P.; Gálvez, Beatriz G.; Arroyo, Alicia G.; Mobashery, Shahriar; Fridman, Rafael.

In: Journal of Biological Chemistry, Vol. 277, No. 29, 19.07.2002, p. 26340-26350.

Research output: Contribution to journalArticle

Toth, M, Hernandez-Barrantes, S, Osenkowski, P, Margarida Bernardo, M, Gervasi, DC, Shimura, Y, Meroueh, O, Kotra, LP, Gálvez, BG, Arroyo, AG, 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, vol. 277, no. 29, pp. 26340-26350. https://doi.org/10.1074/jbc.M200655200
Toth, Marta ; Hernandez-Barrantes, Sonia ; Osenkowski, Pamela ; Margarida Bernardo, M. ; Gervasi, David C. ; Shimura, Yoichiro ; Meroueh, Oussama ; Kotra, Lakshmi P. ; Gálvez, Beatriz G. ; Arroyo, Alicia G. ; Mobashery, Shahriar ; Fridman, Rafael. / Complex pattern of membrane type 1 matrix metalloproteinase shedding. Regulation by autocatalytic cell surface inactivation of active enzyme. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 29. pp. 26340-26350.
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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.",
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AU - Hernandez-Barrantes, Sonia

AU - Osenkowski, Pamela

AU - Margarida Bernardo, M.

AU - Gervasi, David C.

AU - Shimura, Yoichiro

AU - Meroueh, Oussama

AU - Kotra, Lakshmi P.

AU - Gálvez, Beatriz G.

AU - Arroyo, Alicia G.

AU - Mobashery, Shahriar

AU - Fridman, Rafael

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N2 - 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.

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