title: Immunopurification and characterization of a collagenase/gelatinase domain issued from basement membrane fibronectin.
abstract: The proteolytic potential of cellular fibronectin fragments issued from a basement membrane hydrolysate was investigated. Three different gelatinase activities (47, 43 and 37 kDa), located by gelatin zymography, were isolated using successively heparin-agarose, gelatin-agarose and immunopurification with polyclonal antibodies directed against bovine plasma fibronectin. These fragments were also characterized using a monoclonal antibody directed against the extra-domain EDA of cellular fibronectin as a probe. A collagenase activity, reliably indicated by the gelatin zymography pattern, was also found using MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2, the intramolecularly quenched fluorogenic substrate of collagenases. From these results, cellular fibronectin was found to be able to exhibit a proteolytic function after limited proteolysis. This MMP-like function could be associated with tissue remodeling in both normal and pathological states, such as metastasis, angiogenesis and tissue repair.
authors.affiliation: Facultĕ de Médecine Broussais Hôtel-Dieu, Université Pierre et Marie Curie, Paris, France.
authors.forename: L
authors.initials: L
authors.lastname: Boudjennah
authors.forename: V
authors.initials: V
authors.lastname: Dalet-Fumeron
authors.forename: S
authors.initials: S
authors.lastname: Ylätupa
authors.forename: M
authors.initials: M
authors.lastname: Pagano
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001485:Basement Membrane / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D017364:Collagenases / Q000378:metabolism*; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000097:blood / Q000737:chemistry* / Q000378:metabolism*; D018093:Gelatinases / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D000906:Antibodies; D005353:Fibronectins; D009842:Oligopeptides; D010446:Peptide Fragments; D017364:Collagenases; D018093:Gelatinases; D003488:Cyanogen Bromide