title: Characterization of the substrate-binding sites of the mitochondrial nicotinamide nucleotide transhydrogenase.
abstract: The mitochondrial energy-linked nicotinamide nucleotide transhydrogenase (TH) is modified and inhibited by p-fluorosulfonylbenzoyl-5'-adenosine (FSBA). The modification appears to occur at the NAD(H)-binding site when TH alone or TH in the presence of NADPH is incubated with FSBA. However, when this site is protected by NADH, then FSBA inhibits TH more slowly and modifies a different, though specific, site. This second site could be the NADP(H)-binding site. Using [3H]FSBA in the presence of NADPH, the NAD(H)-binding site was modified, and a single tryptic peptide carrying the label was isolated and sequenced. The amino acid sequence of this peptide was Glu-Ser-Gly-Glu-Gly-Gln-Gly-Gly-Tyr*-Ala-Lys. The modified residue was Tyr. The labeled peptide isolated after incubating TH with [3H]FSBA in the presence of NADH could not be completely purified. However, amino acid analysis and partial sequencing made it possible to identify this segment on the amino acid sequence of bovine TH as derived from its cDNA by Yamaguchi et al. (private communication).
authors.affiliation: Department of Basic and Clinical Research, Research Institute of Scripps Clinic, La Jolla, California 92037.
authors.forename: S
authors.initials: S
authors.lastname: Wakabayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Hatefi
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D004926:Escherichia coli / Q000378:metabolism; D066298:In Vitro Techniques; D008929:Mitochondria, Heart / Q000201:enzymology*; D009243:NAD / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D009250:NADP Transhydrogenases / Q000378:metabolism*; D010455:Peptides / Q000032:analysis; D014357:Trypsin
chemical_list: D000596:Amino Acids; D010455:Peptides; D009243:NAD; D009249:NADP; C010959:5'-(4-fluorosulfonylbenzoyl)adenosine; D009247:NADH, NADPH Oxidoreductases; D009250:NADP Transhydrogenases; D014357:Trypsin; D000241:Adenosine