Conformational dynamics of the bovine mitochondrial ADP/ATP carrier isoform 1 revealed by hydrogen/deuterium exchange coupled to mass spectrometry.
The mitochondrial adenine nucleotide carrier (Ancp) catalyzes the transport of ADP and ATP across the mitochondrial inner membrane, thus playing an essential role in cellular energy metabolism. During the transport mechanism the carrier switches between two different conformations that can be blocked by two toxins: carboxyatractyloside (CATR) and bongkrekic acid. Therefore, our understanding of the nucleotide transport mechanism can be improved by analyzing structural differences of the individual inhibited states. We have solved the three-dimensional structure of bovine carrier isoform 1 (bAnc1p) in a complex with CATR, but the structure of the carrier-bongkrekic acid complex, and thus, the detailed mechanism of transport remains unknown. Improvements in sample processing in the hydrogen/deuterium exchange technique coupled to mass spectrometry (HDX-MS) have allowed us to gain novel insights into the conformational changes undergone by bAnc1p. This paper describes the first study of bAnc1p using HDX-MS. Results obtained with the CATR-bAnc1p complex were fully in agreement with published results, thus, validating our approach. On the other hand, the HDX kinetics of the two complexes displays marked differences. The bongkrekic acid-bAnc1p complex exhibits greater accessibility to the solvent on the matrix side, whereas the CATR-bAnc1p complex is more accessible on the intermembrane side. These results are discussed with respect to the structural and biochemical data available on Ancp.
Rey
Martial
M
Laboratoire de Biochimie et Biophysique des Systèmes Intégrés, Institut de Recherches en Technologies et Sciences pour le Vivant, UMR 5092 CNRS-Commissariat à l'Energie Atomique-Université Joseph Fourier, F-38054 Grenoble Cedex 9, France.
Man
Petr
P
Clémençon
Benjamin
B
Trézéguet
Véronique
V
Brandolin
Gérard
G
Forest
Eric
E
Pelosi
Ludovic
L
D000818:Animals; D001278:Atractyloside / Q000031:analogs & derivatives* / Q000737:chemistry; D001692:Biological Transport; D001865:Bongkrekic Acid / Q000737:chemistry*; D002417:Cattle; D041961:Deuterium Exchange Measurement; D007700:Kinetics; D013058:Mass Spectrometry; D000226:Mitochondrial ADP, ATP Translocases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary
D001865:Bongkrekic Acid; D001278:Atractyloside; D000226:Mitochondrial ADP, ATP Translocases; C003853:carboxyatractyloside
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