Further characterization of porcine plasma fibronectin which contains fucosylated carbohydrate chains.
Porcine plasma fibronectin and its functional four fragments produced by cathepsin B digestion were examined for biological, immunochemical and biochemical properties. Native fibronectin, 150-kDa and 130-kDa fragments exhibited similar cell attachment-promoting activity to each other. In an Ouchterlony double immunodiffusion system, these three polypeptides formed a precipitin line with anti-fibronectin antiserum, while the 50-kDa and 30-kDa fragments did not. The 150-kDa and 130-kDa fragments contained free sulfhydryl(s). The glycopeptide fractions were prepared by pronase digestion of porcine and human plasma fibronectin, and radiolabeled with [14C]acetic anhydride. The results of affinity chromatography with concanavalin A and lentil lectin immobilized on agarose indicated that the porcine glycopeptide fraction was different from the human fraction in that a larger part (58%) of the former was bound to lentil lectin. About 90% of this lentil lectin-reactive glycopeptides lost this reactivity upon alpha-L-fucosidase digestion. The glycopeptide fractions were also prepared from three carbohydrate-containing domains. Less than 30% of the radioactivity of the glycopeptide fractions of 150-kDa and 130-kDa fragments was retained on the lentil lectin-agarose, while about 90% of that from the 50-kDa fragment was retained. These results indicate that porcine plasma fibronectin has characteristics very similar to those of human plasma fibronectin and others, but is unique in that it contains fucosylated carbohydrate chains which unevenly distribute through functional domains.
D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002401:Cathepsin B; D002403:Cathepsins / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D003208:Concanavalin A / Q000378:metabolism; D005353:Fibronectins / Q000097:blood* / Q000494:pharmacology; D005779:Immunodiffusion; D037102:Lectins / Q000378:metabolism; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D037121:Plant Lectins; D013552:Swine
D002241:Carbohydrates; D005353:Fibronectins; D037102:Lectins; D010446:Peptide Fragments; D037121:Plant Lectins; C033619:lentil lectin; D003208:Concanavalin A; D002403:Cathepsins; D002401:Cathepsin B