Improved method for rapid purification of protein kinase from streptomycetes.
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.
Janecek
J
J
Institute of Microbiology, Czech Academy of Sciences, Prague, Czech Republic.
Dobrová
Z
Z
Moravec
V
V
Náprstek
J
J
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*
D001426:Bacterial Proteins; D012694:Serine; D012685:Sepharose; D011494:Protein Kinases