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Dimerization of endogenous MT1-MMP is a regulatory step in the activation of the 72-kDa gelatinase MMP-2 on fibroblasts and fibrosarcoma cells

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@article{5dd3bf3c657f45e49b99cc50099baccd,
title = "Dimerization of endogenous MT1-MMP is a regulatory step in the activation of the 72-kDa gelatinase MMP-2 on fibroblasts and fibrosarcoma cells",
abstract = "The secreted gelatinase matrix metalloprotease-2 (MMP-2) and the membrane-anchored matrix metalloprotease MT1-MMP (MMP-14), are central players in pericellular proteolysis in extracellular matrix degradation. In addition to possessing a direct collagenolytic and gelatinolytic activity, these enzymes take part in a cascade pathway in which MT1-MMP activates the MMP-2 proenzyme. This reaction occurs in an interplay with the matrix metalloprotease inhibitor, TIMP-2, and the proposed mechanism involves two molecules of MT1-MMP in complex with one TIMP-2 molecule. We provide positive evidence that proMMP-2 activation is governed by dimerization of MT1-MMP on the surface of fibroblasts and fibrosarcoma cells. Even in the absence of transfection and overexpression, dimerization of MT1-MMP markedly stimulated the formation of active MMP-2 products. The effect demonstrated here was brought about by a monoclonal antibody that binds specifically to MT1-MMP as shown by immunofluorescence experiments. The antibody has no effect on the catalytic activity. The effect on proMMP-2 activation involves MT1-MMP dimerization because it requires the divalent monoclonal antibody, with no effect obtained with monovalent Fab fragments. Since only a negligible level of proMMP-2 activation was obtained with MT1-MMP-expressing cells in the absence of dimerization, our results identify the dimerization event as a critical level of proteolytic cascade regulation.",
keywords = "Animals, Antibodies, Monoclonal, Cattle, Cell Line, Tumor, Dimerization, Enzyme Activation, Enzyme Precursors, Fibroblasts, Hemopexin, Humans, Immunization, Matrix Metalloproteinase 14, Matrix Metalloproteinase 2, Mice, Molecular Weight, Protein Structure, Quaternary, Protein Structure, Tertiary",
author = "Signe Ingvarsen and Madsen, {Daniel H} and Thore Hillig and Lund, {Leif R{\o}ge} and Kenn Holmbeck and Niels Behrendt and Engelholm, {Lars H}",
year = "2008",
month = "7",
doi = "10.1515/BC.2008.097",
language = "English",
volume = "389",
pages = "943--53",
journal = "Biological Chemistry",
issn = "1431-6730",
publisher = "Walter/de Gruyter GmbH & Co. KG",
number = "7",

}

RIS

TY - JOUR

T1 - Dimerization of endogenous MT1-MMP is a regulatory step in the activation of the 72-kDa gelatinase MMP-2 on fibroblasts and fibrosarcoma cells

AU - Ingvarsen, Signe

AU - Madsen, Daniel H

AU - Hillig, Thore

AU - Lund, Leif Røge

AU - Holmbeck, Kenn

AU - Behrendt, Niels

AU - Engelholm, Lars H

PY - 2008/7

Y1 - 2008/7

N2 - The secreted gelatinase matrix metalloprotease-2 (MMP-2) and the membrane-anchored matrix metalloprotease MT1-MMP (MMP-14), are central players in pericellular proteolysis in extracellular matrix degradation. In addition to possessing a direct collagenolytic and gelatinolytic activity, these enzymes take part in a cascade pathway in which MT1-MMP activates the MMP-2 proenzyme. This reaction occurs in an interplay with the matrix metalloprotease inhibitor, TIMP-2, and the proposed mechanism involves two molecules of MT1-MMP in complex with one TIMP-2 molecule. We provide positive evidence that proMMP-2 activation is governed by dimerization of MT1-MMP on the surface of fibroblasts and fibrosarcoma cells. Even in the absence of transfection and overexpression, dimerization of MT1-MMP markedly stimulated the formation of active MMP-2 products. The effect demonstrated here was brought about by a monoclonal antibody that binds specifically to MT1-MMP as shown by immunofluorescence experiments. The antibody has no effect on the catalytic activity. The effect on proMMP-2 activation involves MT1-MMP dimerization because it requires the divalent monoclonal antibody, with no effect obtained with monovalent Fab fragments. Since only a negligible level of proMMP-2 activation was obtained with MT1-MMP-expressing cells in the absence of dimerization, our results identify the dimerization event as a critical level of proteolytic cascade regulation.

AB - The secreted gelatinase matrix metalloprotease-2 (MMP-2) and the membrane-anchored matrix metalloprotease MT1-MMP (MMP-14), are central players in pericellular proteolysis in extracellular matrix degradation. In addition to possessing a direct collagenolytic and gelatinolytic activity, these enzymes take part in a cascade pathway in which MT1-MMP activates the MMP-2 proenzyme. This reaction occurs in an interplay with the matrix metalloprotease inhibitor, TIMP-2, and the proposed mechanism involves two molecules of MT1-MMP in complex with one TIMP-2 molecule. We provide positive evidence that proMMP-2 activation is governed by dimerization of MT1-MMP on the surface of fibroblasts and fibrosarcoma cells. Even in the absence of transfection and overexpression, dimerization of MT1-MMP markedly stimulated the formation of active MMP-2 products. The effect demonstrated here was brought about by a monoclonal antibody that binds specifically to MT1-MMP as shown by immunofluorescence experiments. The antibody has no effect on the catalytic activity. The effect on proMMP-2 activation involves MT1-MMP dimerization because it requires the divalent monoclonal antibody, with no effect obtained with monovalent Fab fragments. Since only a negligible level of proMMP-2 activation was obtained with MT1-MMP-expressing cells in the absence of dimerization, our results identify the dimerization event as a critical level of proteolytic cascade regulation.

KW - Animals

KW - Antibodies, Monoclonal

KW - Cattle

KW - Cell Line, Tumor

KW - Dimerization

KW - Enzyme Activation

KW - Enzyme Precursors

KW - Fibroblasts

KW - Hemopexin

KW - Humans

KW - Immunization

KW - Matrix Metalloproteinase 14

KW - Matrix Metalloproteinase 2

KW - Mice

KW - Molecular Weight

KW - Protein Structure, Quaternary

KW - Protein Structure, Tertiary

U2 - 10.1515/BC.2008.097

DO - 10.1515/BC.2008.097

M3 - Journal article

VL - 389

SP - 943

EP - 953

JO - Biological Chemistry

JF - Biological Chemistry

SN - 1431-6730

IS - 7

ER -

ID: 39988635