Primary structure around the lipoate-attachment site on the E2 component of bovine heart pyruvate dehydrogenase complex.
Bovine heart pyruvate dehydrogenase complex was acetylated by using [3-14C]pyruvate in the presence of N-ethylmaleimide, with approx. 1 mol of acetyl groups being incorporated per mol of E2 polypeptide. After peptic digestion, lipoate-containing peptides were purified by high-voltage electrophoresis and ion-exchange and reverse-phase h.p.l.c. The amino acid sequence around the lipoic acid-attachment site of E2 was determined by automated Edman degradation. Acetylation of a lipoate cofactor bound to a lysine residue was verified by fast-atom-bombardment m.s.
D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D005033:Ethylmaleimide / Q000494:pharmacology; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology*; D010446:Peptide Fragments / Q000032:analysis; D030481:Pyruvate Dehydrogenase (Lipoamide); D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid; D008063:Thioctic Acid / Q000032:analysis*
D010446:Peptide Fragments; D011768:Pyruvate Dehydrogenase Complex; D011773:Pyruvates; D008063:Thioctic Acid; D019289:Pyruvic Acid; D030481:Pyruvate Dehydrogenase (Lipoamide); D005033:Ethylmaleimide