title: Purification of a novel eIF-2 alpha protein kinase from calf brain.
abstract: A new eukaryotic initiation factor 2 kinase has been purified for the first time from calf brain cytosol. The purification of a nonabundant novel protein kinase activity, designated as PKI, that phosphorylates the alpha subunit of eukaryotic initiation factor 2 is described. The protein kinase activity was assayed using purified initiation factor 2 as a substrate and was purified by ammonium sulphate precipitation, conventional chromatography in heparin-Sepharose and phosphocellulose and by high performance size exclusion and anion exchange chromatographies. The protein kinase activity elutes in the region of 140,000 in the size exclusion chromatography and is associated with two different polypeptides a and b, with relative molecular masses of 38,000 and 20,000 and an approximate ratio of 2.5-3.0:1. The protein kinase does not phosphorylate casein or histones and it is independent of cyclic nucleotides. It can be classified as a serine kinase since the phosphorylation of the alpha subunit of eIF-2 is produced in serine residues. Under these conditions none of the kinase subunits are phosphorylated.
authors.affiliation: Dpto. Investigación, Hospital Ramón y Cajal, Madrid, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Alcázar
authors.forename: E
authors.initials: E
authors.lastname: Méndez
authors.forename: M E
authors.initials: ME
authors.lastname: Martín
authors.forename: M
authors.initials: M
authors.lastname: Salinas
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism; D019892:eIF-2 Kinase
chemical_list: D000596:Amino Acids; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D019892:eIF-2 Kinase