title: Involvement of lysine 1047 in type I collagen-mediated activation of polymorphonuclear neutrophils.
abstract: Oxidative functions of polymorphonuclear neutrophils (PMNs), which play a deciding role in the phagocytosis process, are stimulated by extracellular matrix proteins such as type I collagen. Previous studies have demonstrated the involvement of a DGGRYY sequence located within the alpha(1) chain C-terminal telopeptide in type I collagen-induced PMN activation, but so far the mechanism has not been completely elucidated. We have recently demonstrated that collagen carbamylation (i.e. post-translational binding of cyanate to lysine epsilon-NH(2) groups) impairs PMN oxidative functions, suggesting the potential involvement of lysine residues in this process. The present study was devoted to the identification of lysine residues involved in the collagen-induced activation of PMNs. The inhibition of PMN activation by collagen in the presence of 6-amino-hexanoic acid, a structural analogue of lysine residues, confirmed the involvement of specific lysine residues. Modification of lysine residues by carbamylation demonstrated that only one residue, located within the alpha(1)CB6 collagen peptide, was involved in this mechanism. A recombinant alpha(1)CB6 peptide, designed for the substitution of lysine 1047 by glycine, exhibited decreased activity, demonstrating that the lysine residue at position 1047 within the collagen molecule played a significant role in the mechanism of activation. These results help to understand in more detail the collagen-mediated PMN activation mechanism and confirm the prominent involvement of lysine residues in interactions between extracellular matrix proteins and inflammatory cells.
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Faculty of Medicine, University of Reims Champagne-Ardenne, UMR CNRS no. 6237, 51 Rue Cognacq-Jay, Reims, France. stephane.jaisson@univ-reims.fr
authors.forename: Stéphane
authors.initials: S
authors.lastname: Jaisson
authors.forename: Hervé
authors.initials: H
authors.lastname: Sartelet
authors.forename: Corinne
authors.initials: C
authors.lastname: Perreau
authors.forename: Charlotte
authors.initials: C
authors.lastname: Blanchevoye
authors.forename: Roselyne
authors.initials: R
authors.lastname: Garnotel
authors.forename: Philippe
authors.initials: P
authors.lastname: Gillery
mesh_terms: D000595:Amino Acid Sequence; D015119:Aminocaproic Acid / Q000494:pharmacology; D000818:Animals; D024042:Collagen Type I / Q000037:antagonists & inhibitors / Q000737:chemistry*; D003488:Cyanogen Bromide / Q000737:chemistry; D006801:Humans; D008239:Lysine; D008969:Molecular Sequence Data; D018375:Neutrophil Activation / Q000187:drug effects; D009504:Neutrophils / Q000187:drug effects; D010455:Peptides / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D024042:Collagen Type I; D010455:Peptides; D008239:Lysine; D003488:Cyanogen Bromide; D015119:Aminocaproic Acid