Phosphorylation of annexin XI (CAP-50) in SR-3Y1 cells.
Annexin XI (CAP-50) is a probable target protein of calcyclin. Being different from other annexins, annexin XI localizes mainly in nuclei of cultured fibroblasts. In rat embryonic fibroblasts transformed by Rous sarcoma virus oncogene, SR-3Y1 cells, phosphorylation of annexin XI was increased on both serine and threonine residues (Ser < Thr), compared with findings in control 3Y1 cells. The amount of phosphorylated annexin XI was approximately 8.5% of the total cellular annexin XI and the phosphorylated annexin XI migrated slightly slower on SDS-polyacrylamide gel electrophoresis than did the non-phosphorylated form of annexin XI. Phosphorylated annexin XI was recovered in the cytoplasmic fraction and did not bind to phosphatidylserine vesicle in the presence of high Ca2+ (over 1 mM). Annexin XI was phosphorylated by mitogen-activated protein (MAP) kinase, which was reported to be activated in v-src-transformed fibroblast (Gupta, S. K., Gallego, C. Johnson, G.L. and Heasley, L.E. (1992) J. Biol. Chem. 267, 7987-7990), on both serine and threonine residues (Ser >> Thr) in vitro. Comparative phosphopeptide mappings analyzed by reverse-phase high performance liquid chromatography suggested that the sites phosphorylated in situ in SR-3Y1 cells are distinct from the sites by MAP kinase. Annexin XI phosphorylated by MAP kinase still possessed the ability to bind to phosphatidylserine vesicle. These results suggest that annexin XI is a substrate for some Ser/Thr kinase(s) which is activated in v-src-transformed cells and that the phosphorylation may regulate the function of annexin XI in living cells.
Department of Pharmacology, Nagoya University, School of Medicine, Japan.
A
A
Mizutani
H
H
Tokumitsu
R
R
Kobayashi
H
H
Hidaka
D000818:Animals; D017302:Annexins / Q000737:chemistry / Q000378:metabolism*; D001345:Autoradiography; D015153:Blotting, Western; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002458:Cell Fractionation; D002461:Cell Line, Transformed; D016391:Genes, src; D010449:Peptide Mapping; D010743:Phospholipids / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats
D017302:Annexins; D010743:Phospholipids; D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases