Mitochondrial phosphate transport. The Saccharomyces cerevisiae (threonine 43 to cysteine) mutant protein explicitly identifies transport with genomic sequence.
The yeast mitochondrial phosphate transport protein (PTP) has only 38% sequence similarity to the bovine heart protein, and it has recently been postulated to code for a mitochondrial import receptor. Since the reconstitutively active protein is not completely pure, it is important to demonstrate explicitly that the yeast gene codes for PTP. We have replaced Thr43 with Cys (T43C) and show that its unidirectional and pH gradient-dependent inorganic phosphate transport activity becomes highly sensitive to N-ethylmaleimide. This new PTP/T43C catalyzes less than 10% of the wild type transport activity (1 mM [Pi]e, pHe (6.80); 0 mM [Pi]i, pHi (8.07); 30 s [Pi] uptake) suggesting that Thr43 occupies an important position in the PTP.
Phelps
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Boston Biomedical Research Institute, Harvard Medical School, Massachusetts 02114.
Wohlrab
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D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D002384:Catalysis; D003545:Cysteine / Q000235:genetics*; D004247:DNA / Q000235:genetics; D005033:Ethylmaleimide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D028044:Phosphate-Binding Proteins; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013912:Threonine / Q000235:genetics*
D002352:Carrier Proteins; D028044:Phosphate-Binding Proteins; D013912:Threonine; D004247:DNA; D003545:Cysteine; D005033:Ethylmaleimide