Abstract
A new method using hydrophobic interaction chromatography on phenyl-Sepharose was developed to purify branched chain alpha-ketoacid dehydrogenase complex from commercially available frozen rat liver. Yields of greater than 50% were routinely achieved. The purified enzyme, composed of E1 alpha, E1 beta, and E2 subunits, appeared homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and contained endogenous kinase activity for phosphorylation and inactivation of the complex.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)
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Adenosine Triphosphate / pharmacology
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Animals
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Electrophoresis, Polyacrylamide Gel
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Ketone Oxidoreductases / antagonists & inhibitors
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Ketone Oxidoreductases / isolation & purification*
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Ketone Oxidoreductases / metabolism
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Kinetics
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Liver / enzymology*
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Multienzyme Complexes / antagonists & inhibitors
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Multienzyme Complexes / isolation & purification*
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Multienzyme Complexes / metabolism
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Rats
Substances
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Multienzyme Complexes
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Adenosine Triphosphate
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Ketone Oxidoreductases
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3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)