Direct visualization of protease action on collagen triple helical structure

Enzymatic processing of extracellular matrix (ECM) macromolecules by matrix metalloproteases (MMPs) is crucial in mediating physiological and pathological cell processes. However, the molecular mechanisms leading to effective physiological enzyme-ECM interactions remain elusive. Only scant informati...

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Veröffentlicht in:PloS one 2010-06, Vol.5 (6), p.e11043-e11043
Hauptverfasser: Rosenblum, Gabriel, Van den Steen, Philippe E, Cohen, Sidney R, Bitler, Arkady, Brand, David D, Opdenakker, Ghislain, Sagi, Irit
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container_issue 6
container_start_page e11043
container_title PloS one
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creator Rosenblum, Gabriel
Van den Steen, Philippe E
Cohen, Sidney R
Bitler, Arkady
Brand, David D
Opdenakker, Ghislain
Sagi, Irit
description Enzymatic processing of extracellular matrix (ECM) macromolecules by matrix metalloproteases (MMPs) is crucial in mediating physiological and pathological cell processes. However, the molecular mechanisms leading to effective physiological enzyme-ECM interactions remain elusive. Only scant information is available on the mode by which matrix proteases degrade ECM substrates. An example is the enzymatic degradation of triple helical collagen II fragments, generated by the collagenase MMP-8 cleavage, during the course of acute inflammatory conditions by gelatinase B/MMP-9. As is the case for many other matrix proteases, it is not clear how MMP-9 recognizes, binds and digests collagen in this important physiological process. We used single molecule imaging to directly visualize this protease during its interaction with collagen fragments. We show that the initial binding is mediated by the diffusion of the protease along the ordered helix on the collagen (3/4) fragment, with preferential binding of the collagen tail. As the reaction progressed and prior to collagen degradation, gelatin-like morphologies resulting from the denaturation of the triple helical collagen were observed. Remarkably, this activity was independent of enzyme proteolysis and was accompanied by significant conformational changes of the working protease. Here we provide the first direct visualization of highly complex mechanisms of macromolecular interactions governing the enzymatic processing of ECM substrates by physiological protease.
doi_str_mv 10.1371/journal.pone.0011043
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As the reaction progressed and prior to collagen degradation, gelatin-like morphologies resulting from the denaturation of the triple helical collagen were observed. Remarkably, this activity was independent of enzyme proteolysis and was accompanied by significant conformational changes of the working protease. Here we provide the first direct visualization of highly complex mechanisms of macromolecular interactions governing the enzymatic processing of ECM substrates by physiological protease.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>20585385</pmid><doi>10.1371/journal.pone.0011043</doi><tpages>e11043</tpages><oa>free_for_read</oa></addata></record>
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subjects Binding
Bioavailability
Biochemistry
Biochemistry/Macromolecular Assemblies and Machines
Biophysics/Macromolecular Assemblies and Machines
Collagen
Collagen (type II)
Collagen - chemistry
Collagen - metabolism
Collagenase
Cytokines
Degradation
Denaturation
Electrophoresis, Polyacrylamide Gel
Enzymes
Extracellular matrix
Extracellular Matrix - metabolism
Fragmentation
Fragments
Gelatin
Gelatinase
Gelatinase B
Inflammation
Laboratories
Macromolecules
Matrix metalloproteinases
Matrix Metalloproteinases - metabolism
Medical research
Microscopy, Atomic Force
Molecular modelling
Neutrophil collagenase
Neutrophils
Peptides
Physiological aspects
Physiology
Protease
Proteases
Protein Conformation
Protein denaturation
Proteinase
Proteins
Proteolysis
Science
Substrates
Visualization
Yoga
title Direct visualization of protease action on collagen triple helical structure
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