Subunit Structure of Dihydrolipoamide Acetyltransferase Component of Pyruvate Dehydrogenase Complex from Bovine Kidney

Abstract

The pyruvate dehydrogenase complex was isolated from bovine kidney mitochondria under special anti-proteolytic conditions yielding preparations with a specific activity of up to 20 U[units]/mg protein. Dihydrolipoamide acetyltransferase resolved from the complex was subjected to limited proteolysis resulting in the formation of 2 major fragments with apparent MW of 36,000 and 28,000. The fragments were isolated by extraction from dodecyl sulfate polyacrylamide gels and possessed enzymatic activity for acetyl transfer. Acetylation studies indicated that each fragment contains 1 protein-bound lipoyl group. The kidney dihydrolipoamide acetyltransferase subunit consists of 2 homologous if not identical domains. A model is suggested where the acetyltransferase core of the mammalian pyruvate dehydrogenase complex is made up of 30 polypeptide chains whose 60 domains could be arranged in pentagonal dodecahedron symmetry quite similar as proposed for the 60 subunit structure of the acetyltransferase core.