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Localizing Ligand Binding Sites Using Overlapping Recombinant Polypeptide Sequences Of Vitronectin, Jodi L. Watson

Masters Theses

Vitronectin is a glycoprotein involved in many cellular processes including blood coagulation, fibrinolysis, and cell/matrix binding. It interacts with a number of macromolecules including heparin, PAI-1, and the thrombin-antithrombin complex. We have studied the interaction of full-length vitronectin and the thrombin-antithrombin complex using a Far Western method. To localize the recognition site for thrombin-antithrombin binding to vitronectin, sequences for eight overlapping recombinant polypeptide sequences of vitronectin (spanning all 459 amino acids) were cloned into the pET system. Expression in BL21(DE3)pLysS Escherichia coli has been achieved for seven of the eight 100 amino acid fragments. Seven polypeptides (amino acids 1-100, 51-150, …