Improved method for rapid purification of protein kinase from streptomycetes
Language English Country Netherlands Media print
Document type Journal Article
PubMed
8926341
DOI
10.1016/0165-022x(95)00022-j
PII: 0165-022X(95)00022-J
Knihovny.cz E-resources
- MeSH
- Bacterial Proteins isolation & purification MeSH
- Chromatography, Affinity methods MeSH
- Protein Kinases isolation & purification MeSH
- Sepharose MeSH
- Serine MeSH
- Streptomyces enzymology MeSH
- Chromatography, High Pressure Liquid MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Bacterial Proteins MeSH
- Protein Kinases MeSH
- Sepharose MeSH
- Serine MeSH
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.
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