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Phosphate transport and proteins with SH groups in rat liver mitochondria
Authors:S Alziari  S Touraille  Y Briand  R Durand
Institution:Laboratoire de Biochimie, Université de Clermont II, B.P. 45, 63170 Aubière, France
Abstract:Phosphate transport in rat liver mitochondria was studied by following 32P] phosphate uptake within physiological concentrations. Transport inhibition due to mersalyl and protection by mersalyl against N-ethylmaleimide measured in those conditions corresponded to earlier results obtained by the swelling technique. When mitochondria were incubated with 3H] N-ethylmaleimide in the presence of mersalyl, the radioactive labeling in proteins of particles obtained after sonication was decreased in all fractions, but three proteins were both highly alkylated and also highly protected by mersalyl (M.W. 48,000 - 36,000 - 31,000). Two of these (M.W. 36,000 and 31,000) were partially purified by ultrogel chromatography in the presence of sodium dodecyl sulfate. Furthermore, it was shown that both phosphate and nigericin diminished labeling by N-ethylmaleimide in the final supernatant fraction. Two proteins (M.W. 98,000 and 31,000) were significantly alkylated by 3H] N-ethylmaleimide and protected by phosphate and nigericin.
Keywords:mitochondria  phosphate transport  NEM  N-ethylmaleimide  Tris  Tris (hydroxy methyl) aminomethane  HEPES  N-2-hydroxyethyl piperazine-N′-2 ethane sulfonic acid  EGTA  Ethyleneglycol-bis (β amino ethyl ether) tetraacetic acid  PPO  2  5-diphenyloxazole  POPOP  1  4-di(2-(4-methyl-5-phenyloxazolyl))  SDS  Sodium dodecyl sulfate  Pi  Inorganic phosphate  ClCCP  Chlorocarbonyl cyanide phenyl hydrazone
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