Conformational changes of the yeast mitochondrial adenosine diphosphate/adenosine triphosphate carrier studied through its intrinsic fluorescence. 1. Tryptophanyl residues of the carrier can be mutated without impairing protein activity

Biochemistry
A Le SauxGuy J-M Lauquin

Abstract

During the transport process the mitochondrial adenine nucleotide carrier (Ancp) undergoes conformational changes which result in modifications of the intrinsic fluorescence of the carrier. To further study these changes by a fluorometric approach, the three tryptophanyl residues (Trp87, Trp126, and Trp235) of the Saccharomyces cerevisiae Anc2p were individually mutated to their tyrosine counterparts. The resulting mutated genes (two-Trp, one-Trp or Trp-less variants) were integrated at the ANC2 locus. A prerequisite for such studies is that all the engineered carrier molecules are still able to catalyze ADP/ATP exchange. The cellular characteristics of the strains expressing the mutated Anc2p and the biochemical properties of the variant Anc2p in mitochondria were examined. Although Trp87 is absolutely conserved in all 30 available Ancp sequences, none of the tryptophanyl residues is essential to the carrier protein folding and the transport activity. The mutated and wild-type Anc2p were expressed to the same level, as evidenced by both ligand binding and immunochemical analyses. When isolated in the presence of detergent, all the variant Anc2p preparations contained ergosterol in similar amounts (9 mol/mol of 35 kDa Anc2p) bu...Continue Reading

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Citations

Nov 4, 2008·Journal of Bioenergetics and Biomembranes·Véronique TrézéguetGérard Brandolin
Aug 27, 2011·The Journal of Biological Chemistry·Benjamin ClémençonLudovic Pelosi
Feb 3, 2006·Annual Review of Biochemistry·H NuryE Pebay-Peyroula
May 10, 2011·Biochimie·Amandine ThomasLudovic Pelosi
Feb 29, 2000·Biochimica Et Biophysica Acta·V TrézéguetG J Lauquin
Mar 16, 2005·Protein Expression and Purification·Emmanuel Philippe DassaGérard Brandolin
May 31, 2008·Biochimica Et Biophysica Acta·Martin Klingenberg
May 31, 2016·Biochimie·Benjamin ClémençonMatthias A Hediger
Mar 13, 2012·International Journal of Molecular Sciences·Benjamin Clémençon
Feb 27, 2008·The Journal of Biological Chemistry·Benjamin ClémençonLudovic Pelosi
Nov 9, 2005·Biochemistry·Vincent PostisVéronique Trézéguet

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