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Relations structure-fonction des transporteurs nucléotides

Abstract : Chloroplast NTT1 mediates external supply of ATP in the plastids, which is pre- requisite for the maintenance of plastid metabolism during limited or missing photosynthetic activity. However, their molecular mechanism remains poorly understood, primarily due to the difficulty of producing and purifying functional recombinant forms of these transporters. We have successfully developed a protocol for the production of this transporter, that is compatible for efficient solubilization and good yield of purified protein. Combining biochemical and biophysical analyses, we could characterize the protein solution and identify conditions from which first crystals could be obtained. To further extend our work with nucleotide transporter, structural and functional studies on mitochondrial ADP/ATP carrier (AAC) have been performed. AAC and NTT1 belong to different families but exhibit similar functional features. To discover small-molecule inhibitors of AAC's transport for structural studies, virtual docking of compounds was performed into the AAC active site, using the already known structure of AAC. Docking tools were installed and screening a large library of 75,000 compounds allowed the identification of 17 molecules. The experimental binding assay for AAC, revealed that one of the compounds has inhibitory activity in the micromolar regime. In addition, 3 novel ADP analogues showing inhibitory effect against ATP transport of AAC have also been identified.
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Submitted on : Tuesday, April 3, 2012 - 3:31:47 PM
Last modification on : Saturday, March 26, 2022 - 3:17:28 AM
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  • HAL Id : tel-00684264, version 1



Pankaj Panwar. Relations structure-fonction des transporteurs nucléotides. Sciences agricoles. Université de Grenoble, 2012. Français. ⟨NNT : 2012GRENV005⟩. ⟨tel-00684264⟩



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