title: Improved method for rapid purification of protein kinase from streptomycetes.
abstract: Protein kinase from Streptomyces lincolnensis was purified nearly to homogeneity using a high performance liquid chromatography (HPLC) and a Pharmacia FPLC system. The procedure used employed column chromatography on DE-53, followed by FPLC affinity chromatography with serine- or threonine-Sepharose (prepared as described in this paper) and gel filtration using a Superose 12 or TSK G3000SW column. Starting with 3.5 g of mycelial proteins, approximately 1 mg of pure enzyme was obtained. The procedure is simple and highly reproducible. The protein kinase thus obtained was nearly pure by silver staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified protein kinase phosphorylated substrate proteins at the seryl residues.
authors.affiliation: Institute of Microbiology, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Janecek
authors.forename: Z
authors.initials: Z
authors.lastname: Dobrová
authors.forename: V
authors.initials: V
authors.lastname: Moravec
authors.forename: J
authors.initials: J
authors.lastname: Náprstek
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D011494:Protein Kinases / Q000302:isolation & purification*; D012685:Sepharose; D012694:Serine; D013302:Streptomyces / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D012694:Serine; D012685:Sepharose; D011494:Protein Kinases