Laboratoire de Chimie Bactérienne UMR 7283

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16 novembre 2017: 1 événement


  • Séminaire IMM

    Jeudi 16 novembre 10:30-12:00 - Gilles Truan - INSA Toulouse

    Séminaire BIP

    Résumé : Séminaire ouvert à tous !

    Analysis of the large conformational changes of the NADPH cytochrome P450 reductase by incorporation of non natural amino acids, fluorophore labeling and FRET techniques.
    Résumé :

    NADPH cytochrome P450 reductase (CPR) is a multidomain protein that transfers electrons from NADPH to a series of electron acceptors. CPR comprises one FAM and one FAD domains that contains the flavins, responsible for the various electrons transfer steps. Large conformational changes have been evidenced in CPR. They trigger the movements of the FMN and FAD domains that are mandatory to promote electron transfers to acceptors. We have demonstrated that CPR is actually in a rapid conformational equilibrium between two states. We recently produced CPR having incorporated two different unnatural amino acids (cyclopropene-L-lysine and p-propargyloxy-L-phenylalanine) in the two catalytic domains. These amino acids were directly derivatized with Cy3 and Cy5, thus producing CPR molecules bearing two different fluorophores. The talk will be centered on the development of the technique as well as the preliminary results obtained in single molecule microscopy FRET.

    Lieu : Salle Jacques Senez

    En savoir plus : Séminaire IMM