Conformational dependence of collagenase (matrix metalloproteinase-1) up-regulation by elastin peptides in cultured fibroblasts

J Biol Chem. 2001 Feb 16;276(7):5222-7. doi: 10.1074/jbc.M003642200. Epub 2000 Nov 17.

Abstract

We have established that treatment of cultured human skin fibroblasts with tropoelastin or with heterogenic peptides, obtained after organo-alkaline or leukocyte elastase hydrolysis of insoluble elastin, induces a high expression of pro-collagenase-1 (pro-matrix metalloproteinase-1 (pro-MMP-1)). The identical effect was achieved after stimulation with a VGVAPG synthetic peptide, reflecting the elastin-derived domain known to bind to the 67-kDa elastin-binding protein. This clearly indicated involvement of this receptor in the described phenomenon. This notion was further reinforced by the fact that elastin peptides-dependent MMP-1 up-regulation has not been demonstrated in cultures preincubated with 1 mm lactose, which causes shedding of the elastin-binding protein and with pertussis toxin, which blocks the elastin-binding protein-dependent signaling pathway involving G protein, phospholipase C, and protein kinase C. Moreover, we demonstrated that diverse peptides maintaining GXXPG sequences can also induce similar cellular effects as a "principal" VGVAPG ligand of the elastin receptor. Results of our biophysical studies suggest that this peculiar consensus sequence stabilizes a type VIII beta-turn in several similar, but not identical, peptides that maintain a sufficient conformation to be recognized by the elastin receptor. We have also established that GXXPG elastin-derived peptides, in addition to pro-MMP-1, cause up-regulation of pro-matrix metalloproteinase-3 (pro-stromelysin 1). Furthermore, we found that the presence of plasmin in the culture medium activated these MMP proenzymes, leading to a consequent degradation of collagen substrate. Our results may be, therefore, relevant to pathobiology of inflammation, in which elastin-derived peptides bearing the GXXPG conformation (created after leukocyte-dependent proteolysis) bind to the elastin receptor of local fibroblasts and trigger signals leading to expression and activation of MMP-1 and MMP-3, which in turn exacerbate local connective tissue damage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Circular Dichroism
  • Collagen / metabolism
  • Collagenases / biosynthesis
  • Collagenases / genetics*
  • Collagenases / metabolism
  • Consensus Sequence
  • Elastin / chemistry*
  • Elastin / pharmacology*
  • Enzyme Precursors / biosynthesis
  • Enzyme Precursors / genetics*
  • Enzyme Precursors / metabolism
  • Fibroblasts / metabolism*
  • Humans
  • Matrix Metalloproteinase 1
  • Metalloendopeptidases / biosynthesis
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism
  • Peptides / chemistry
  • Peptides / pharmacology
  • Protein Conformation
  • RNA, Messenger / biosynthesis
  • Receptors, Cell Surface / physiology
  • Up-Regulation*

Substances

  • Enzyme Precursors
  • Peptides
  • RNA, Messenger
  • Receptors, Cell Surface
  • elastin-binding proteins
  • Collagen
  • Elastin
  • Collagenases
  • Metalloendopeptidases
  • prostromelysin
  • Matrix Metalloproteinase 1