Cloning, Expression, and Type II Collagenolytic Activity of Matrix metalloproteinase-13 From Human Osteoarthritic Cartilage

J Clin Invest. 1996 Feb 1;97(3):761-8. doi: 10.1172/JCI118475.

Abstract

Proteolysis of triple-helical collagen is an important step in the progression toward irreversible tissue damage in osteoarthritis. Earlier work on the expression of enzymes in cartilage suggested that collagenase-1 (MMP-1) contributes to the process. Degenerate reverse transcription polymerase chain reaction experiments, Northern blot analysis, and direct immunodetection have now provided evidence that collagenase-3 (MMP-13), an enzyme recently cloned from human breast carcinoma, is expressed by chondrocytes in human osteoarthritic cartilage. Variable levels of MMP-13 and MMP-1 in cartilage was significantly induced at both the message and protein levels by interleukin-1 alpha. Recombinant MMP-13 cleaved type II collagen to give characteristic 3/4 and 1/4 fragments; however, MMP-13 turned over type II collagen at least 10 times faster than MMP-1. Experiments with intact type II collagen as well as a synthetic peptide suggested that MMP-13 cleaved type II collagen at the same bond as MMP-1, but this was then followed by a secondary cleavage that removed three amino acids from the 1/4 fragment amino terminus. The expression of MMP-13 in osteoarthritic cartilage and its activity against type II collagen suggest that the enzyme plays a significant role in cartilage collagen degradation, and must consequently form part of a complex target for proposed therapeutic interventions based on collagenase inhibition.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cartilage / enzymology*
  • Cloning, Molecular
  • Collagen / metabolism*
  • Collagenases / genetics
  • Collagenases / metabolism*
  • Humans
  • Kinetics
  • Matrix Metalloproteinase 13
  • Molecular Sequence Data
  • Osteoarthritis / enzymology*
  • Polymerase Chain Reaction
  • Sequence Analysis
  • Sequence Homology, Amino Acid
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Substrate Specificity

Substances

  • Collagen
  • Collagenases
  • MMP13 protein, human
  • Matrix Metalloproteinase 13

Associated data

  • SWISSPROT/P02458