Full-length and truncated forms of vitronectin provide insight into effects of proteolytic processing on function.
A genetic polymorphism in the vitronectin allele directs the production of two distinct forms of the 459 amino acid glycoprotein. A methionine present at position 381 favors production of the single-chain form of vitronectin, while threonine at this position increases the susceptibility of vitronectin to cleavage just beyond its heparin-binding domain at residue 379. This reaction gives rise to a disulfide-bonded, two-chain form of vitronectin. In order to investigate the functional significance of the vitronectin polymorphism, the baculovirus system has been used to express recombinant full-length vitronectin and a truncated form of the molecule that represents the 62-kDa fragment of two-chain vitronectin. Both forms of vitronectin bind and neutralize heparin anticoagulant activity. The proteins also bind PAI-1 and stabilize its active conformation. These experiments suggest that the C-terminal 80 amino acids do not confer a functional difference in the two allelic variants. Immunoassays and gel filtration experiments indicate that both full-length and truncated recombinant forms of vitronectin are multimeric. Together with other reports from this laboratory, these results provide information regarding the primary binding sites for two vitronectin ligands and further define regions that may be involved in multimerization of the protein.
Gibson
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M407 Walters Life Sciences Building, Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA.
Peterson
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D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics; D006493:Heparin / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D017924:Nucleopolyhedroviruses / Q000235:genetics; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D011487:Protein Conformation / Q000187:drug effects; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D017384:Sequence Deletion; D018411:Spodoptera / Q000166:cytology; D013329:Structure-Activity Relationship; D013917:Thrombin / Q000037:antagonists & inhibitors; D019096:Vitronectin / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D046911:Macromolecular Substances; D017395:Plasminogen Activator Inhibitor 1; D011993:Recombinant Fusion Proteins; D019096:Vitronectin; D006493:Heparin; D013917:Thrombin