A new method using hydrophobic interaction chromatography on phenyl-Sepharose was developed to purify branched chain α-ketoacid dehydrogenase complex from commercially available frozen rat liver. Yields of greater than 50% were routinely achieved. The purified enzyme, composed of E1α, E1β, and E2 subunits, appeared homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and contained endogenous kinase activity for phosphorylation and inactivation of the complex.
- branched chain α-ketoacid dehydrogenase complex
- branched chain α-ketoacid dehydrogenase kinase
- hydrophobic interaction chromatography
- rat liver
ASJC Scopus subject areas
- Molecular Biology