High-Chloride Concentrations Abolish the Binding of Adenine Nucleotides in the Mitochondrial ADP/ATP Carrier Family

Krammer, Eva-Maria; Ravaud, Stéphanie; Dagognet, François; Frelet-Barrand, Annie; Pebay‐Peyroula, Eva; Chipot, Christophe

doi:10.1016/j.bpj.2009.08.047

Cited by 20 publications

(21 citation statements)

References 16 publications

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“…A report [44] indicates that sensory rhodopsin has a binding site for Cl -, which regulates rhodopsin activity. Further, it has been reported that Cl -regulates mitochondrial ADP/ATP carrier by inhibiting the binding of adenine nucleotides to the carrier [45], and that ENaC activity in the kidney is regulated intracellular Cl - [33]. These reports and our observations in the present study suggest that Cl -plays important roles in cell functions.…”

Section: Discussionsupporting

confidence: 80%

Chloride Ion Modulates Cell Proliferation of Human Androgen-independent Prostatic Cancer Cell

Hiraoka

Mitsuya

Niisato

et al. 2010

Cell Physiol Biochem

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In the present study, we investigated if the intracellular Cl^- affects cell growth and cell cycle progression of androgen-independent prostate cancer PC3 cells. PC3 cells cultured in a medium containing 113 mM Cl^- for 96 h grew up 9-fold in cell number, while PC3 cells cultured in an 8 mM-Cl^--containing culture medium showed complete arrest of cell growth even after culture for 96 h. Exposure of cells to the 8 mM-Cl^- culture medium diminished phosphorylation levels of Rb and cdc2, which are respectively key accelerators of transition from G₁ to S phase and G₂ to M phase in cell cycle progression. Culturing cells in the 8 mM-Cl^--containing culture medium upregulated the protein expression level of p21 (a CDK inhibitor) inhibiting transition of G₁ to S phase, and diminished the incorporation of 5-ethynyl-2′-deoxyuridine (EdU; a thymidine analogue) into DNA. These results suggest that cells cultured in the low Cl^- medium prolonged the duration of all phases of the cell cycle (G₁, S, and G₂/M), thereby abolishing overall cell cycle progression. Effects of culturing cells in the low Cl^- culture medium on cell cycle progression would be mediated via a change in the intracellular Cl^- concentration ([Cl^-]_i), since [Cl^-]_i was decreased under a low Cl^- culture medium. To clarify this possibility, we studied effects of furosemide and bumetanide, Na⁺/K⁺/2Cl^- cotransporter (NKCC) inhibitors, on proliferation of PC3 cells. Furosemide and bumetanide decreased [Cl^-]_i and cell growth of PC3 cells. These results suggest that a change in [Cl^-]_i would play a critical role in this growth mechanism.

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Section: Discussionsupporting

confidence: 80%

Chloride Ion Modulates Cell Proliferation of Human Androgen-independent Prostatic Cancer Cell

Hiraoka

Mitsuya

Niisato

et al. 2010

Cell Physiol Biochem

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“…In its crystallographic structure, AAC is in a conformation open toward the inter-membrane space, fully occluded on the matrix side and impervious to water or any other small substrate 19,21,44 . The MD simulations performed here enlighten, however, striking differences in the structure of the two proteins.…”

Section:  Discussionmentioning

confidence: 99%

Dangerous Liaisons between Detergents and Membrane Proteins. The Case of Mitochondrial Uncoupling Protein 2

et al. 2013

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The extraction of membrane proteins from their native environment by detergents is central to their biophysical characterization. Recent studies have emphasized that detergents may perturb the structure locally and modify the dynamics of membrane proteins. However, it remains challenging to determine whether these perturbations are negligible or could be responsible for misfolded conformations, altering the protein's function. In this work, we propose an original strategy combining functional studies and molecular simulations to address the physiological relevance of membrane protein structures obtained in the presence of detergents. We apply our strategy to a structure of isoform 2 of an uncoupling protein (UCP2) binding an inhibitor recently obtained in dodecylphosphocholine detergent micelles. Although this structure shares common traits with the ADP/ATP carrier, a member of the same protein family, its functional and biological significance remains to be addressed. In the present investigation, we demonstrate how dodecylphosphocholine severely alters the structure as well as the function of UCPs. The proposed original strategy opens new vistas for probing the physiological relevance of three-dimensional structures of membrane proteins obtained in non-native environments.

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“…The positively charged carrier residues, which electrostatically interact with the phosphate groups, drive the passage of the substrate through the cavity to its bottom [21,22]. In fact, high chloride concentration conditions cause inhibition of binding and the further transport of nucleotides [23]. Moreover, tyrosine residues facing the cavity are the targets of SRC kinases that modulate transport by phosphorylation [24].…”

Section: Introductionmentioning

confidence: 99%

Mapping multiple potential ATP binding sites on the matrix side of the bovine ADP/ATP carrier by the combined use of MD simulation and docking

et al. 2011

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The mitochondrial adenosine diphosphate/adenosine triphosphate (ADP/ATP) carrier-AAC-was crystallized in complex with its specific inhibitor carboxyatractyloside (CATR). The protein consists of a six-transmembrane helix bundle that defines the nucleotide translocation pathway, which is closed towards the matrix side due to sharp kinks in the odd-numbered helices. In this paper, we describe the interaction between the matrix side of the AAC transporter and the ATP(4-) molecule using carrier structures obtained through classical molecular dynamics simulation (MD) and a protein-ligand docking procedure. Fifteen structures were extracted from a previously published MD trajectory through clustering analysis, and 50 docking runs were carried out for each carrier conformation, for a total of 750 runs ("MD docking"). The results were compared to those from 750 docking runs performed on the X-ray structure ("X docking"). The docking procedure indicated the presence of a single interaction site in the X-ray structure that was conserved in the structures extracted from the MD trajectory. MD docking showed the presence of a second binding site that was not found in the X docking. The interaction strategy between the AAC transporter and the ATP(4-) molecule was analyzed by investigating the composition and 3D arrangement of the interaction pockets, together with the orientations of the substrate inside them. A relationship between sequence repeats and the ATP(4-) binding sites in the AAC carrier structure is proposed.

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High-Chloride Concentrations Abolish the Binding of Adenine Nucleotides in the Mitochondrial ADP/ATP Carrier Family

Cited by 20 publications

References 16 publications

Chloride Ion Modulates Cell Proliferation of Human Androgen-independent Prostatic Cancer Cell

Chloride Ion Modulates Cell Proliferation of Human Androgen-independent Prostatic Cancer Cell

Dangerous Liaisons between Detergents and Membrane Proteins. The Case of Mitochondrial Uncoupling Protein 2

Mapping multiple potential ATP binding sites on the matrix side of the bovine ADP/ATP carrier by the combined use of MD simulation and docking

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