title: Conformational dependence of collagenase (matrix metalloproteinase-1) up-regulation by elastin peptides in cultured fibroblasts.
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.
authors.affiliation: UPRES-A CNRS 6021, IFR53 Biomolécules, Faculties of Sciences and Medicine, IFR53 Biomolécules, Faculty of Sciences, University of Reims, 51687 Reims, France.
authors.forename: B
authors.initials: B
authors.lastname: Brassart
authors.forename: P
authors.initials: P
authors.lastname: Fuchs
authors.forename: E
authors.initials: E
authors.lastname: Huet
authors.forename: A J
authors.initials: AJ
authors.lastname: Alix
authors.forename: J
authors.initials: J
authors.lastname: Wallach
authors.forename: A M
authors.initials: AM
authors.lastname: Tamburro
authors.forename: F
authors.initials: F
authors.lastname: Delacoux
authors.forename: B
authors.initials: B
authors.lastname: Haye
authors.forename: H
authors.initials: H
authors.lastname: Emonard
authors.forename: W
authors.initials: W
authors.lastname: Hornebeck
authors.forename: L
authors.initials: L
authors.lastname: Debelle
mesh_terms: D002478:Cells, Cultured; D002942:Circular Dichroism; D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D016384:Consensus Sequence; D004549:Elastin / Q000737:chemistry* / Q000494:pharmacology*; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D020781:Matrix Metalloproteinase 1; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D011487:Protein Conformation; D012333:RNA, Messenger / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000502:physiology; D015854:Up-Regulation
chemical_list: D004792:Enzyme Precursors; D010455:Peptides; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; C054901:elastin-binding proteins; D003094:Collagen; D004549:Elastin; D017364:Collagenases; D008666:Metalloendopeptidases; C048047:prostromelysin; D020781:Matrix Metalloproteinase 1