Neurospora crassa pyruvate dehydrogenase complex: component characterization, catalytic properties and location of translation.
We propose a simplified procedure for the purification of the Neurospora crassa pyruvate dehydrogenase complex. The purified complex showed four protein bands with apparent Mr values of 53,400, 52,900, 49,000 and 36,900 upon SDS-polyacrylamide gel electrophoresis. Components, E2 and E3, of N. crassa pyruvate dehydrogenase complex were identified, respectively, as polypeptides 49,000 and 53,400. It can be deduced that component E1 is constituted of two subunits with Mr values of 52,900 and 36,900. The Km values towards different substrates and the optimal pH and temperature were determined. The protein kinase activity associated with the core enzyme was present in our most highly purified preparations. It was demonstrated that all the protein components of the complex are synthesized under the control of the nuclear genome.
Mareck
A M
AM
Laboratoire de Biochimie, Unité Associée au CNRS No. 203, Faculté des Sciences, Université de Rouen Haute-Normandie, Mont-Saint-Aignan, France.
Bessam
H
H
Foucher
B
B
D002118:Calcium / Q000494:pharmacology; D002701:Chloramphenicol / Q000494:pharmacology; D003065:Coenzyme A / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*; D014176:Protein Biosynthesis / Q000187:drug effects; D011768:Pyruvate Dehydrogenase Complex / Q000302:isolation & purification / Q000378:metabolism*; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid
D011768:Pyruvate Dehydrogenase Complex; D011773:Pyruvates; D009243:NAD; D002701:Chloramphenicol; D019289:Pyruvic Acid; D003513:Cycloheximide; D008274:Magnesium; D005033:Ethylmaleimide; D003065:Coenzyme A; D002118:Calcium