TY - JOUR

T1 - MMP-12 catalytic domain recognizes and cleaves at multiple sites in human skin collagen type I and type III

AU - Taddese, Samuel

AU - Jung, Michael C

AU - Ihling, Christian

AU - Heinz, Andrea

AU - Neubert, Reinhard H H

AU - Schmelzer, Christian E H

N1 - Copyright 2009 Elsevier B.V. All rights reserved.

PY - 2010/4

Y1 - 2010/4

N2 - Collagens of either soft connective or mineralized tissues are subject to continuous remodeling and turnover. Undesired cleavage can be the result of an imbalance between proteases and their inhibitors. Owing to their superhelical structure, collagens are resistant to many proteases and matrix metalloproteinases (MMPs) are required to initiate further degradation by other enzymes. Several MMPs are known to degrade collagens, but the action of MMP-12 has not yet been studied in detail. In this work, the potential of MMP-12 in recognizing sites in human skin collagen types I and III has been investigated. The catalytic domain of MMP-12 binds to the triple helix and cleaves the typical sites -Gly(775)-Leu(776)- in alpha-2 type I collagen and -Gly(775)-Ile(776)- in alpha-1 type I and type III collagens and at multiple other sites in both collagen types. Moreover, it was observed that the region around these typical sites contains comparatively less prolines, of which some have been proven to be only partially hydroxylated. This is of relevance since partial hydroxylation in the vicinity of a potential scissile bond may have a local effect on the conformational thermodynamics with probable consequences on the collagenolysis process. Taken together, the results of the present work confirm that the catalytic domain of MMP-12 alone binds and degrades collagens I and III.

AB - Collagens of either soft connective or mineralized tissues are subject to continuous remodeling and turnover. Undesired cleavage can be the result of an imbalance between proteases and their inhibitors. Owing to their superhelical structure, collagens are resistant to many proteases and matrix metalloproteinases (MMPs) are required to initiate further degradation by other enzymes. Several MMPs are known to degrade collagens, but the action of MMP-12 has not yet been studied in detail. In this work, the potential of MMP-12 in recognizing sites in human skin collagen types I and III has been investigated. The catalytic domain of MMP-12 binds to the triple helix and cleaves the typical sites -Gly(775)-Leu(776)- in alpha-2 type I collagen and -Gly(775)-Ile(776)- in alpha-1 type I and type III collagens and at multiple other sites in both collagen types. Moreover, it was observed that the region around these typical sites contains comparatively less prolines, of which some have been proven to be only partially hydroxylated. This is of relevance since partial hydroxylation in the vicinity of a potential scissile bond may have a local effect on the conformational thermodynamics with probable consequences on the collagenolysis process. Taken together, the results of the present work confirm that the catalytic domain of MMP-12 alone binds and degrades collagens I and III.

KW - Amino Acid Sequence

KW - Binding Sites

KW - Catalytic Domain

KW - Chromatography, High Pressure Liquid

KW - Collagen

KW - Collagen Type I

KW - Collagen Type III

KW - Humans

KW - Hydroxylation

KW - In Vitro Techniques

KW - Matrix Metalloproteinase 12

KW - Molecular Sequence Data

KW - Peptide Fragments

KW - Protein Conformation

KW - Protein Structure, Tertiary

KW - Recombinant Proteins

KW - Skin

KW - Spectrometry, Mass, Electrospray Ionization

KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

KW - Tandem Mass Spectrometry

KW - Thermodynamics

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1016/j.bbapap.2009.11.014

DO - 10.1016/j.bbapap.2009.11.014

M3 - Journal article

C2 - 19932771

VL - 1804

SP - 731

EP - 739

JO - Biochimica et Biophysica Acta - Reviews on Cancer

JF - Biochimica et Biophysica Acta - Reviews on Cancer

SN - 0304-419X

IS - 4

ER -