Agonist-induced phosphorylation of an immunologically ras-related protein in human erythroleukemia cells.
Monoclonal antibody M90 recognizes a specific epitope of the ras-encoded p21 protein. This region comprises amino acids 107-130 containing the residues 116-119, which are related to GTP binding. This antibody strongly reacts on Western Blots with a 22kDa protein from human erythroleukemia (HEL) cells. Treatment of HEL cells with iloprost, an agonist that increases cellular cyclic AMP levels, produces the appearance of a protein with an apparent molecular mass of 24kDa. This protein is also recognized by antiserum M90 on Western Blots; its appearance parallels a decrease of the 22kDa protein, and it can be labeled with 32P. This effect is also observed with dibutyryl cyclic AMP, which indicates phosphorylation of the 22kDa protein by cyclic AMP-dependent protein kinase. This phosphorylation produces an electrophoretic mobility change of the 22kDa protein to a 24kDa region on gels. The change of mobility of the 22kDa protein induced by iloprost in HEL cells is also observed when the protein is labeled with [35S]methionine and immunoprecipitated with antiserum M90. This information indicates a coupling mechanism involving phosphorylation of an oncogene product in HEL cells.
Lazarowski
E R
ER
Division of Cell Biology, Burroughs Wellcome Co., Research Triangle Park, N.C. 27709.
Lacal
J C
JC
Lapetina
E G
EG
D000911:Antibodies, Monoclonal; D003994:Bucladesine / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D000939:Epitopes / Q000032:analysis; D011464:Epoprostenol / Q000494:pharmacology; D006801:Humans; D016285:Iloprost; D004915:Leukemia, Erythroblastic, Acute; D008565:Membrane Proteins / Q000378:metabolism*; D008970:Molecular Weight; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D016283:Proto-Oncogene Proteins p21(ras); D013806:Theophylline / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D003994:Bucladesine; D013806:Theophylline; D011464:Epoprostenol; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras); D016285:Iloprost