Improved method for rapid purification of protein kinase from streptomycetes
Jazyk angličtina Země Nizozemsko Médium print
Typ dokumentu časopisecké články
PubMed
8926341
DOI
10.1016/0165-022x(95)00022-j
PII: 0165-022X(95)00022-J
Knihovny.cz E-zdroje
- MeSH
- bakteriální proteiny izolace a purifikace MeSH
- chromatografie afinitní metody MeSH
- proteinkinasy izolace a purifikace MeSH
- sefarosa MeSH
- serin MeSH
- Streptomyces enzymologie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bakteriální proteiny MeSH
- proteinkinasy MeSH
- sefarosa MeSH
- serin 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.
Citace poskytuje Crossref.org
General and molecular microbiology and microbial genetics in the IM CAS
