Cyclic AMP-dependent protein kinase phosphorylates serine378 in vitronectin.
We previously observed that Ser378 in the heparin-binding domain of vitronectin becomes phosphorylated by a protein kinase in plasma upon addition of ATP and divalent cations. We now report that purified plasma vitronectin contains approximately 2.5 mol of phosphate per mol of protein and that vitronectin becomes phosphorylated during biosynthesis in human hepatoma (HepG2) cells. In vitro, rabbit muscle cAMP-dependent protein kinase specifically phosphorylates Ser378 in single-chain (75 kDa) vitronectin but does not phosphorylate the two-chain (65/10 kDa) form cleaved at Arg379. Heparin affects neither the time course nor the extent of phosphorylation of Ser378 at neutral pH. The extent of phosphorylation of Ser378 achieved with cAMP-dependent protein kinase (greater than or equal to 0.3 mol phosphate per mol vitronectin) is greater than that obtainable in plasma and should enable comparisons to be made of the activities of the native and phosphorylated forms.
Mehringer
J H
JH
Department of Internal Medicine, Washington University, St. Louis, Missouri 63110.
Weigel
C J
CJ
Tollefsen
D M
DM
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000378:metabolism; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D010450:Endopeptidases; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008113:Liver Neoplasms; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D010449:Peptide Mapping; D010710:Phosphates / Q000378:metabolism; D010748:Phosphopeptides / Q000302:isolation & purification; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D012694:Serine; D013379:Substrate Specificity; D019096:Vitronectin
D001798:Blood Proteins; D006023:Glycoproteins; D010446:Peptide Fragments; D010710:Phosphates; D010748:Phosphopeptides; D019096:Vitronectin; D012694:Serine; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D010450:Endopeptidases