Complete primary structure of the collagen-binding domain of bovine fibronectin

Abstract

The complete amino acid sequence of the collagen-binding domain of bovine plasma fibronectin has been determined. The fragment, generated by digestion of fibronectin with plasmin and chymotrypsin, contains 340 residues (260 -599 of fibronectin) with threonine and tryptophan as the amino-terminal and carboxyl-terminal amino acids, respectively. 24 half-cystines and no cysteines are present in the sequence. Three glucosamine-based oligosaccharide groups are attached to Asn-399, Asn-497 and to Asn-511, respectively. Two of the three types (I and 11) [Petersen et al. (1983) Proc. Natl Acad. Sci. U S A 80,137 -1411 of internal homology occur in the fragment, namely four of the at least twelve stretches of type I sequence homology, 'fingers', and two stretches of type I1 homology. The type I homology is present in two other plasmic fragments from fibronectin, while the type I1 homology has been found in the collagen-binding domain only.Fibronectin is a multiple-domain glycoprotein ( M , ~4 5 0 0 0 0 ) composed of two identical or nearly identical polypeptide chains, linked by two disulfide bridges near the C terminus [I]. It is found on the surface of many different cell types [2] and in blood plasma [3]. Its manifold functions are associated with cell adhesion, cell spreading and motility, opsonization, wound healing and the maintenance of normal morphology (reviewed in [4-71) In addition it can bind to both vertebrate and bacterial cell surfaces [16, 171. As a result of a reaction catalyzed by factor XIII, (blood plasma transglutaminase), fibronectin can become covalently cross-linked to collagen [18], fibrin [9, 191 and to the surface of Staphylococcus aureus [20].Approximately one-half of the amino acid sequence of bovine plasma fibronectin has been determined [I], and three types of internal homology (types I -111) recognized [l]. Specific limited proteolysis of bovine plasma fibronectin with plasmin releases four major fragments with M, of 29000, 170000,23000 and 6000 [21], mentioned in the order they occur from the N terminus of fibronectin. The complete amino acid sequence of the 29-kDa, 23-kDa and 6-kDa fragments have been determined [I], and several cyanogen bromide (CNBr) fragments, originating from the 170-kDa fragment have also been sequenced [I, 221. The 29-kDa fragment (259 residues) consists almost entirely of five mutually homologous type I domains ('fingers') [I]. The 23-kDa fragment (178 residues) contains another three type I domains [I], and has recently been overlapped to the 6-kDa fragment [23]. The latter is a dimer of two identical 26-residue peptides linked to each other by two disulfide bridges (the interchain bridges) [I]; it contains a serine-0-phosphate near the C terminus [22]. Limited proAbbreviations. HPLC, high-performance liquid chromatography; Hse, homoserine; HVE, high-voltage paper electrophoresis; SDS, sodium dodecyl sulfate; dansyl, 5-dimethylaminonaphthalene-I-sulfonyl.Enzymes. Chymotrypsin (EC 3.4.21.1); trypsin (EC 3.4.21.4); plasmin (EC 3.4.21.7); Staphylococcus aureus V8 p...

Isolation and characterization of gelatin-binding proteins from goat seminal plasma

“…In addition, they contain the characteristic sequence found in type II domains [21], particularly the motif CVFPFXY(R/K)X(R/K)(R/K)(H/Y)F that is much conserved among BSPs and other homologs found in boar and stallion seminal plasma proteins [22]. In GSP-20 kDa and GSP-22 kDa, the motif is not strictly respected, markedly the FPF part replaced by F A F, but the general pattern of amino acids is followed.…”

Crystallization and preliminary X-ray diffraction analysis of bovine seminal plasma PDC-109, a protein composed of two fibronectin type II domains

“…1 Each of these PDC-109 domains displays the invariant residues and the topology of the disulfide bonds of the fibronectin type II (Fn II) module. 2 The occurrence of fibronectin type II-like domains is relatively widespread. The Fn II module is present as a single copy in blood coagulation factor XIII (Hageman factor), 3 the mannose receptor, 4 the insulinlike growth factor II receptor (cation-independent mannose-6-phosphate receptor), 5,6 the DEC-205 receptor, 7 and the hepatocyte growth factor activator.…”

The collagen‐binding site of type‐II units of bovine seminal fluid protein PDC‐109 and fibronectin

“…The two type-I1 units of fibronectin are found in the collagen-binding region of the molecule and have long been suspected to be involved in collagen binding (for a recent review on fibronectin see Ruoslahti, 1988). The collagen-binding site of fibronectin, however, could be only roughly delineated by limited proteolysis of the molecule; the smallest collagen-binding fragments were found to contain, beside the two type-I1 homologous units, a variable number of type-I domains (Skorstengaard et al, 1984). A finer mapping of the collagen-binding site has been performed recently by the analysis of the gelatin-Sepharose affinity of a series of fibronectin fragments expressed in E. coli as P-galactosidase fusion proteins .…”