MMP-20 is predominately a tooth-specific enzyme with a deep catalytic pocket that hydrolyzes type V collagen

Biochemistry. 2006 Mar 28;45(12):3863-74. doi: 10.1021/bi052252o.

Abstract

Matrix metalloproteinase-20 (MMP-20, enamelysin) has a highly restricted pattern of expression. In healthy tissues, MMP-20 is observed in the enamel organ and pulp organ of developing teeth and is present only as an activated enzyme. To identify other tissues that may express MMP-20, we performed a systematic mouse tissue expression screen. Among the non-tooth tissues assayed, MMP-20 transcripts were identified only in minute quantities within the large intestine. The murine Mmp20 promoter was cloned, sequenced, and assessed for potential tooth-specific regulatory elements. In silico analysis identified four promoter modules that were common to Mmp20 and at least two of three coregulated predominantly tooth-specific genes that encode ameloblastin, amelogenin, and enamelin. We asked if the highly restricted MMP-20 expression pattern was associated with a broad substrate specificity that might preclude its expression in other tissues. An iterative mixture-based random doedecamer peptide library screen with Edman sequencing of MMP-20 cleavage products revealed that, among MMPs previously screened, MMP-20 had unique substrate preferences. These preferences indicate that MMP-20 has a deep and wide catalytic pocket that can accommodate substrates with large aromatic residues in the P1' position. On the basis of matrices derived from the peptide library data, we identified and then confirmed that type V collagen is an MMP-20 substrate. Since type V collagen is not present in dental enamel but is an otherwise widely distributed collagen, and since only active MMP-20 has been observed in teeth, our data suggest that control of MMP-20 activity is primarily regulated by transcriptional means.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Base Sequence
  • Blotting, Southern
  • Catalytic Domain
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Collagen Type V / metabolism*
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Hydrolysis
  • Matrix Metalloproteinase 20
  • Matrix Metalloproteinases / chemistry
  • Matrix Metalloproteinases / genetics
  • Matrix Metalloproteinases / isolation & purification
  • Matrix Metalloproteinases / metabolism*
  • Mice
  • Promoter Regions, Genetic
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tooth / enzymology*

Substances

  • Collagen Type V
  • DNA Primers
  • Recombinant Proteins
  • Matrix Metalloproteinase 20
  • Matrix Metalloproteinases
  • Mmp20 protein, mouse

Grants and funding