Purification of branched chain α-ketoacid dehydrogenase complex from rat liver

Yoshiharu Shimomura, Ralph Paxton, Takayuki Ozawa, Robert Harris

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)74-78
Number of pages5
JournalAnalytical Biochemistry
Volume163
Issue number1
DOIs
StatePublished - May 15 1987

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3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)
Phosphorylation
Chromatography
Electrophoresis
Hydrophobic and Hydrophilic Interactions
Sodium Dodecyl Sulfate
Liver
Purification
Rats
Polyacrylamide Gel Electrophoresis
Phosphotransferases
Enzymes
polyacrylamide gels
phenyl-sepharose

Keywords

  • branched chain α-ketoacid dehydrogenase complex
  • branched chain α-ketoacid dehydrogenase kinase
  • hydrophobic interaction chromatography
  • rat liver

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Purification of branched chain α-ketoacid dehydrogenase complex from rat liver. / Shimomura, Yoshiharu; Paxton, Ralph; Ozawa, Takayuki; Harris, Robert.

In: Analytical Biochemistry, Vol. 163, No. 1, 15.05.1987, p. 74-78.

Research output: Contribution to journalArticle

Shimomura, Yoshiharu ; Paxton, Ralph ; Ozawa, Takayuki ; Harris, Robert. / Purification of branched chain α-ketoacid dehydrogenase complex from rat liver. In: Analytical Biochemistry. 1987 ; Vol. 163, No. 1. pp. 74-78.
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