Isolation of a cDNA clone for the dihydrolipoamide acetyltransferase component of the human liver pyruvate dehydrogenase complex.
Dihydrolipoamide acetyltransferase (E2) forms the structural core of pyruvate dehydrogenase complex. A cDNA clone (lambda E2-1) for mammalian E2 was identified from a human liver lambda gt11 library using anti-E2 serum. Affinity-selected antibodies using the fusion protein from lambda E2-1 immuno-reacted specifically with E2 of purified pyruvate dehydrogenase complex on immuno-blot analysis. The cDNA insert was approximately 2.3 kb in length with an internal EcoR1 site generating 1.4 and 0.9 kb fragments. A synthetic 17-mer oligodeoxynucleotide mixture based on the amino acid sequence surrounding the lipoic acid-containing lysine residue in bovine kidney E2 hybridized with the 2.3 kb cDNA insert and the 1.4 kb fragment.
D000123:Acetyltransferases / Q000235:genetics*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI; D051047:Dihydrolipoyllysine-Residue Acetyltransferase; D006801:Humans; D007163:Immunosorbent Techniques; D008930:Mitochondria, Liver / Q000201:enzymology; D011768:Pyruvate Dehydrogenase Complex / Q000032:analysis*
D011768:Pyruvate Dehydrogenase Complex; D004247:DNA; D000123:Acetyltransferases; D051047:Dihydrolipoyllysine-Residue Acetyltransferase; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI