title: Isolation and characterisation of the pyruvate dehydrogenase complex of anaerobically grown Enterococcus faecalis NCTC 775.
abstract: In this contribution the isolation and some of the structural and kinetic properties of the pyruvate dehydrogenase complex (PDC) of anaerobically grown Enterococcus faecalis are described. The complex closely resembles the PDC of other Gram-positive bacteria and eukaryotes. It consists of four polypeptide chains with apparent molecular masses on SDS/PAGE of 97, 55, 42 and 36 kDa, and these polypeptides could be assigned to dihydrolipoyl transacetylase (E2), lipoamide dehydrogenase (E3) and the two subunits of pyruvate dehydrogenase (E1 alpha and E1 beta), respectively. The E2 core has an icosahedral symmetry. The apparent molecular mass on SDS/PAGE of 97 kDa of the E2 chain is extremely high in comparison with other Gram-positive organisms (and eukaryotes) and probably due to several lipoyl domains associated with the E2 chain. NADH inhibition is mediated via E3. The mechanism of inhibition is discussed in view of the high PDC activities in vivo that are found in E. faecalis, grown under anaerobic conditions.
authors.affiliation: Department of Microbiology, Biotechnology Centre, University of Amsterdam, The Netherlands.
authors.forename: J L
authors.initials: JL
authors.lastname: Snoep
authors.forename: A H
authors.initials: AH
authors.lastname: Westphal
authors.forename: J A
authors.initials: JA
authors.lastname: Benen
authors.forename: M J
authors.initials: MJ
authors.lastname: Teixeira de Mattos
authors.forename: O M
authors.initials: OM
authors.lastname: Neijssel
authors.forename: A
authors.initials: A
authors.lastname: de Kok
mesh_terms: D000123:Acetyltransferases / Q000378:metabolism; D002384:Catalysis; D002474:Cell-Free System; D008058:Dihydrolipoamide Dehydrogenase / Q000378:metabolism; D051047:Dihydrolipoyllysine-Residue Acetyltransferase; D013293:Enterococcus faecalis / Q000201:enzymology* / Q000254:growth & development; D006868:Hydrolysis; D007700:Kinetics; D008854:Microscopy, Electron; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D011768:Pyruvate Dehydrogenase Complex / Q000302:isolation & purification* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D011768:Pyruvate Dehydrogenase Complex; D009243:NAD; D008058:Dihydrolipoamide Dehydrogenase; D000123:Acetyltransferases; D051047:Dihydrolipoyllysine-Residue Acetyltransferase