An enzymatic assay for vitronectin based on its selective phosphorylation by protein kinase A.
The catalytic subunit (C) of cAMP-dependent protein kinase selectively phosphorylates vitronectin, a plasma protein that promotes cell adhesion and platelet aggregation, inhibits the inactivation of thrombin by antithrombin III, and participates in complement function. This specific phosphorylation is used here (a) to develop an enzymatic assay for vitronectin (with C and [gamma-32P]ATP) which can be used to identify the vitronectin-containing fractions at each stage of its purification; (b) to radioactively label vitronectin and differentiate between the intact and the nicked form of this protein in structure-function studies; and (c) to identify possible vitronectin-related proteins in the plasma of other animal species.
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
B
B
Korc-Grodzicki
D
D
Chain
T
T
Kreizman
S
S
Shaltiel
D000818:Animals; D001798:Blood Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D006168:Guinea Pigs; D006801:Humans; D051379:Mice; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D013379:Substrate Specificity; D019096:Vitronectin
D001798:Blood Proteins; D006023:Glycoproteins; D019096:Vitronectin; D011494:Protein Kinases