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Defining Requirements for Collagenase Cleavage in Collagen Type III Using a Bacterial Collagen System

Degradation of fibrillar collagens is important in many physiological and pathological events. These collagens are resistant to most proteases due to the tightly packed triple-helical structure, but are readily cleaved at a specific site by collagenases, selected members of the matrix metalloprotein...

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Autores principales: Yu, Zhuoxin, Visse, Robert, Inouye, Masayori, Nagase, Hideaki, Brodsky, Barbara
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3391134/
https://www.ncbi.nlm.nih.gov/pubmed/22573319
http://dx.doi.org/10.1074/jbc.M112.348979
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author Yu, Zhuoxin
Visse, Robert
Inouye, Masayori
Nagase, Hideaki
Brodsky, Barbara
author_facet Yu, Zhuoxin
Visse, Robert
Inouye, Masayori
Nagase, Hideaki
Brodsky, Barbara
author_sort Yu, Zhuoxin
collection PubMed
description Degradation of fibrillar collagens is important in many physiological and pathological events. These collagens are resistant to most proteases due to the tightly packed triple-helical structure, but are readily cleaved at a specific site by collagenases, selected members of the matrix metalloproteinases (MMPs). To investigate the structural requirements for collagenolysis, varying numbers of GXY triplets from human type III collagen around the collagenase cleavage site were inserted between two triple helix domains of the Scl2 bacterial collagen protein. The original bacterial CL domain was not cleaved by MMP-1 (collagenase 1) or MMP-13 (collagenase 3). The minimum type III sequence necessary for cleavage by the two collagenases was 5 GXY triplets, including 4 residues before and 11 residues after the cleavage site (P4-P11′). Cleavage of these chimeric substrates was not achieved by the catalytic domain of MMP-1 or MMP-13, nor by full-length MMP-3. Kinetic analysis of the chimeras indicated that the rate of cleavage by MMP-1 of the chimera containing six triplets (P7-P11′) of collagen III was similar to that of native collagen III. The collagenase-susceptible chimeras were cleaved very slowly by trypsin, a property also seen for native collagen III, supporting a local structural relaxation of the triple helix near the collagenase cleavage site. The recombinant bacterial-human collagen system characterized here is a good model to investigate the specificity and mechanism of action of collagenases.
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spelling pubmed-33911342012-07-11 Defining Requirements for Collagenase Cleavage in Collagen Type III Using a Bacterial Collagen System Yu, Zhuoxin Visse, Robert Inouye, Masayori Nagase, Hideaki Brodsky, Barbara J Biol Chem Protein Structure and Folding Degradation of fibrillar collagens is important in many physiological and pathological events. These collagens are resistant to most proteases due to the tightly packed triple-helical structure, but are readily cleaved at a specific site by collagenases, selected members of the matrix metalloproteinases (MMPs). To investigate the structural requirements for collagenolysis, varying numbers of GXY triplets from human type III collagen around the collagenase cleavage site were inserted between two triple helix domains of the Scl2 bacterial collagen protein. The original bacterial CL domain was not cleaved by MMP-1 (collagenase 1) or MMP-13 (collagenase 3). The minimum type III sequence necessary for cleavage by the two collagenases was 5 GXY triplets, including 4 residues before and 11 residues after the cleavage site (P4-P11′). Cleavage of these chimeric substrates was not achieved by the catalytic domain of MMP-1 or MMP-13, nor by full-length MMP-3. Kinetic analysis of the chimeras indicated that the rate of cleavage by MMP-1 of the chimera containing six triplets (P7-P11′) of collagen III was similar to that of native collagen III. The collagenase-susceptible chimeras were cleaved very slowly by trypsin, a property also seen for native collagen III, supporting a local structural relaxation of the triple helix near the collagenase cleavage site. The recombinant bacterial-human collagen system characterized here is a good model to investigate the specificity and mechanism of action of collagenases. American Society for Biochemistry and Molecular Biology 2012-06-29 2012-05-09 /pmc/articles/PMC3391134/ /pubmed/22573319 http://dx.doi.org/10.1074/jbc.M112.348979 Text en © 2012 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) applies to Author Choice Articles
spellingShingle Protein Structure and Folding
Yu, Zhuoxin
Visse, Robert
Inouye, Masayori
Nagase, Hideaki
Brodsky, Barbara
Defining Requirements for Collagenase Cleavage in Collagen Type III Using a Bacterial Collagen System
title Defining Requirements for Collagenase Cleavage in Collagen Type III Using a Bacterial Collagen System
title_full Defining Requirements for Collagenase Cleavage in Collagen Type III Using a Bacterial Collagen System
title_fullStr Defining Requirements for Collagenase Cleavage in Collagen Type III Using a Bacterial Collagen System
title_full_unstemmed Defining Requirements for Collagenase Cleavage in Collagen Type III Using a Bacterial Collagen System
title_short Defining Requirements for Collagenase Cleavage in Collagen Type III Using a Bacterial Collagen System
title_sort defining requirements for collagenase cleavage in collagen type iii using a bacterial collagen system
topic Protein Structure and Folding
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3391134/
https://www.ncbi.nlm.nih.gov/pubmed/22573319
http://dx.doi.org/10.1074/jbc.M112.348979
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