Yeast ADP/ATP carrier isoform 2: conformational dynamics and role of the RRRMMM signature sequence methionines

Autor: Clémençon, Benjamin, Rey, Martial, Trézéguet, Véronique, Forest, Eric, Pelosi, Ludovic
Přispěvatelé: Laboratoire de Biochimie et Biophysique des Systèmes Intégrés (lBBSI), Université Joseph Fourier - Grenoble 1 (UJF), Institut de biochimie et génétique cellulaires (IBGC), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie de Masse des Protéines (LSMP), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Jazyk: angličtina
Rok vydání: 2011
Předmět:
Saccharomyces cerevisiae Proteins
MESH: Mitochondria
Amino Acid Motifs
Biological Transport
Active
Saccharomyces cerevisiae
Atractyloside
MESH: Protein Isoforms
MESH: Amino Acid Motifs
Methionine
MESH: Saccharomyces cerevisiae Proteins
Membrane Biology
MESH: Anti-Bacterial Agents
Protein Isoforms
[SDV.BBM]Life Sciences [q-bio]/Biochemistry
Molecular Biology
MESH: Bongkrekic Acid
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Structural Biology [q-bio.BM]
MESH: Transcription Factor TFIID
Deuterium Exchange Measurement
MESH: Saccharomyces cerevisiae
Anti-Bacterial Agents
Mitochondria
MESH: Atractyloside
MESH: Deuterium Exchange Measurement
MESH: Methionine
MESH: Biological Transport
Active
Transcription Factor TFIID
MESH: Mitochondrial ADP
ATP Translocases
Bongkrekic Acid
Mitochondrial ADP
ATP Translocases
Zdroj: Journal of Biological Chemistry
Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2011, 286 (41), pp.36119-36131. ⟨10.1074/jbc.M111.277376⟩
Journal of Biological Chemistry, 2011, 286 (41), pp.36119-36131. ⟨10.1074/jbc.M111.277376⟩
ISSN: 0021-9258
1083-351X
DOI: 10.1074/jbc.M111.277376⟩
Popis: International audience; The mitochondrial ADP/ATP carrier, or Ancp, is a member of the mitochondrial carrier family responsible for exchanging ADP and ATP across the mitochondrial inner membrane. ADP/ATP transport involves Ancp switching between two conformational states. These can be analyzed using specific inhibitors, carboxyatractyloside (CATR) and bongkrekic acid (BA). The high resolution three-dimensional structure of bovine Anc1p (bAnc1p), as a CATR-carrier complex, has been solved. However, because the structure of the BA-carrier complex has not yet been determined, the detailed mechanism of transport remains unknown. Recently, sample processing for hydrogen/deuterium exchange experiments coupled to mass spectrometry was improved, providing novel insights into bAnc1p conformational transitions due to inhibitor binding. In this work we performed both hydrogen/deuterium exchange-mass spectrometry experiments and genetic manipulations. Because these are very difficult to apply with bovine Anc1p, we used Saccharomyces cerevisiae Anc isoform 2 (ScAnc2p). Significant differences in solvent accessibility were observed throughout the amino acid sequence for ScAnc2p complexed to either CATR or BA. Interestingly, in detergent solution, the conformational dynamics of ScAnc2p were dissimilar to those of bAnc1p, in particular for the upper half of the cavity, toward the intermembrane space, and the m2 loop, which is thought to be easily accessible to the solvent from the matrix in bAnc1p. Our study then focused on the methionyl residues of the Ancp signature sequence, RRRMMM. All our results indicate that the methionine cluster is involved in the ADP/ATP transport mechanism and confirm that the Ancp cavity is a highly dynamic structure.
Databáze: OpenAIRE
Externí odkaz: