title: Characterization of soluble fibronectin binding to Bacille Calmette-Guérin.
abstract: Fibronectin (FN), a 420 kDa glycoprotein, consists of two similar subunits linked by a disulphide bond near the C-terminal end. FN is present in soluble and matrix forms in various body fluids and tissues and has been shown to bind to variety of organisms. We characterized the conditions required for 125I-FN binding to Bacille Calmette-Guérin (BCG). The binding was dose-dependent, reached saturation within 3 min, and was essentially irreversible for at least 24 h under optimal binding conditions at pH 6.0. In contrast, the binding was reversible (greater than 90% in 24 h) when the pH was increased to 10.0. Scatchard analysis of the dose-response experiments produced a straight line, suggesting the presence of a single class of FN receptor on BCG. 125I-FN binding was trypsin-sensitive, suggesting that the BCG-binding molecule is a protein. The number of FN receptors was determined to be 8000-15,000 per bacterium. 125I-FN binding was pH dependent, with maximal binding at acidic pH. 125I-FN binding was sensitive to the presence of NaCl, with 0.08 M-NaCl inhibiting binding by 85%. These data demonstrate that soluble FN binds to a trypsin-sensitive cell-surface component of BCG in an essentially irreversible manner.
authors.affiliation: Washington University School of Medicine, St. Louis, Missouri.
authors.forename: J
authors.initials: J
authors.lastname: Aslanzadeh
authors.forename: E J
authors.initials: EJ
authors.lastname: Brown
authors.forename: S P
authors.initials: SP
authors.lastname: Quillin
authors.forename: J K
authors.initials: JK
authors.lastname: Ritchey
authors.forename: T L
authors.initials: TL
authors.lastname: Ratliff
mesh_terms: D005353:Fibronectins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009163:Mycobacterium bovis / Q000378:metabolism*; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic / Q000378:metabolism; D012965:Sodium Chloride; D012995:Solubility
chemical_list: D005353:Fibronectins; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic; D012965:Sodium Chloride