title: Characterization of rainbow trout branched-chain alpha-keto acid dehydrogenase complex: inter-domain segments of the E2 component affect the overall activity.
abstract: Branched-chain alpha-keto acid dehydrogenase complex (BCKADH) contains decarboxylase (E1), dihydrolipoyl transacylase (E2), and dihydrolipoyl dehydrogenase (E3) as catalytic components. BCKADH purified from rainbow trout (Oncorhynchus mykiss) liver was comparable with mammalian BCKADH in various enzymatic characteristics, but less efficient in catalyzing the overall reaction. The trout E2 subunit was larger than the mammalian subunit and rather similar to the chicken one in relative molecular mass on SDS-PAGE, whereas the E1 component was similar between trout and mammalian both in relative molecular mass of its alpha and beta subunits and in the catalytic activity. Trout E2 cDNA cloning and nucleotide sequencing revealed that the mature trout E2 subunit consists of 435 residues, and possesses 14 additional residues compared with mammalian E2. Eleven of these are localized in two interdomain segments as two sequences with two and nine residues, respectively. Trout E2 was inferior to rat E2 in the capacity for binding the E1 component, similar to chicken E2. Thus, it appears that non-mammalian BCKADH E2 is distinct from that in mammals in the structure of interdomain segments, resulting in reduction of overall activity of the enzyme complex.
authors.affiliation: Department of Biochemistry, Fukushima Medical University, School of Medicine, 1 Hikarigaoka, Fukushima 960-1295, Japan.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Hakozaki
authors.forename: Kazuo
authors.initials: K
authors.lastname: Ono
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Suzuki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Hata
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Mori
authors.forename: Hideo
authors.initials: H
authors.lastname: Kochi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D002645:Chickens; D003001:Cloning, Molecular; D007655:Ketoglutarate Dehydrogenase Complex / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000235:genetics; D017434:Protein Structure, Tertiary; D021122:Protein Subunits; D051381:Rats; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D021122:Protein Subunits; D007655:Ketoglutarate Dehydrogenase Complex