title: A single mutation in uncoupling protein of rat brown adipose tissue mitochondria abolishes GDP sensitivity of H+ transport.
abstract: The uncoupling protein is one of a family of mitochondrial transport proteins involved in energy metabolism. It dissipates oxidative energy to generate heat, either by catalyzing proton transport directly or by catalyzing fatty acid anion transport, thus enabling fatty acids to act as cycling protonophores. This transport process is tightly regulated by purine nucleotides. We have expressed uncoupling protein in yeast and examined its proton transport activity after its reconstitution into proteoliposomes. A directed change of Arg276 to Leu or Gln completely abolished nucleotide inhibition of protonophoretic action of the reconstituted mutant uncoupling proteins without affecting the transport process. Arg276 is the first residue of functional importance to be identified in uncoupling protein. Mutation of the homologous residue in the yeast ADP/ATP translocator prevented the growth of yeast on a nonfermentable carbon source, presumably by interfering with nucleotide exchange (Nelson, D. R., Lawson, J. E., Klingenberg, M., and Douglas, M. G. (1993) J. Mol. Biol. 230, 1159-1170). Demonstration of the essential role of a single homologous residue in protein-nucleotide interaction within these two transporters is the first direct evidence that uncoupling protein and the ADP/ATP translocator belong to the same gene family.
authors.affiliation: Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
authors.forename: D L
authors.initials: DL
authors.lastname: Murdza-Inglis
authors.forename: M
authors.initials: M
authors.lastname: Modriansky
authors.forename: H V
authors.initials: HV
authors.lastname: Patel
authors.forename: G
authors.initials: G
authors.lastname: Woldegiorgis
authors.forename: K B
authors.initials: KB
authors.lastname: Freeman
authors.forename: K D
authors.initials: KD
authors.lastname: Garlid
mesh_terms: D002001:Adipose Tissue, Brown / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000235:genetics; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D003545:Cysteine / Q000235:genetics; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006859:Hydrogen / Q000378:metabolism*; D007473:Ion Channels; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D024101:Mitochondrial Proteins; D009154:Mutation; D010641:Phenotype; D051381:Rats; D011994:Recombinant Proteins; D000071256:Uncoupling Protein 1
chemical_list: D002352:Carrier Proteins; D007473:Ion Channels; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D011994:Recombinant Proteins; D000071256:Uncoupling Protein 1; D006153:Guanosine Diphosphate; D006859:Hydrogen; D001120:Arginine; D003545:Cysteine