Characterization of the dihydrolipoamide acetyltransferase of the mitochondrial pyruvate dehydrogenase complex from potato and comparisons with similar enzymes in diverse plant species.
The pyruvate dehydrogenase complex (mPDC) from potato (Solanum tuberosum cv. Romano) can be disassociated in 1 M NaCl and 0.1 M glycine into a large dihydrolipoamide acetyltransferase (E2) complex and smaller pyruvate dehydrogenase (E1) and dihydrolipoamide dehydrogenase (E3) complexes. The E2 complex consists of 55 and 78-kDa polypeptides which are reversibly radiolabelled to a similar degree in the intact mPDC by [2-14C]pyruvate. Affinity-purified antibodies against the 55-kDa protein do not cross-react with the 78-kDa protein and the two proteins show different peptide patterns following partial proteolysis. The 78 and 55-kDa proteins are present in approximately equal abundance in the E2 complex and incorporate a similar amount of [14C] on incubation with [2-14C]pyruvate. Native mPDC and the E2 complex have sedimentation coefficients of 50S and 30S, respectively. Titration of electro-eluted polypeptides against the intact mPDC and E2 complex revealed that each mg of mPDC contains 0.4 mg of E1, 0.4 mg of E2 and 0.2 mg of E3. Labelling of partially purified mPDC from potato, pea, cauliflower, maize and barley, with [2-14C]pyruvate, suggest that a 78-kDa acetylatable protein is only found in the dicotyledonous species, while all plant species tested contained a smaller 52-60 kDa acetylatable protein.
Millar
A H
AH
Department of Plant Sciences, University of Oxford, UK. Harvey.Millar@plants.ox.ac.uk
Leaver
C J
CJ
Hill
S A
SA
D000107:Acetylation; D000123:Acetyltransferases / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D000906:Antibodies; D051047:Dihydrolipoyllysine-Residue Acetyltransferase; D007120:Immunochemistry; D046911:Macromolecular Substances; D008928:Mitochondria / Q000201:enzymology; D008970:Molecular Weight; D010944:Plants / Q000201:enzymology*; D011487:Protein Conformation; D011768:Pyruvate Dehydrogenase Complex / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D011817:Rabbits; D011198:Solanum tuberosum / Q000201:enzymology*; D013045:Species Specificity
D000906:Antibodies; D046911:Macromolecular Substances; D011768:Pyruvate Dehydrogenase Complex; D000123:Acetyltransferases; D051047:Dihydrolipoyllysine-Residue Acetyltransferase