Ammonia channeling in bacterial glucosamine-6-phosphate synthase (Glms): molecular dynamics simulations and kinetic studies of protein mutants

Abstract : Ammonia transfer from the glutamine site to the fructose-6P site of bacterial glucosamine-6-phosphate synthase was studied by molecular dynamics simulations. The studies suggest a key role for Trp74, in the sealing of the hydrophobic channel connecting the two binding sites, as well as for the two Ala602 and Val605 residues, which form a narrow passage whose opening/closing constitutes an essential event in ammonia transfer. Kinetic analyses of the corresponding protein mutants confirmed our predictions. The efficiency of ammonia transfer which was close to zero in the W74A mutant was partially restored by increasing the size of the corresponding side-chain; the simulations performed on the W74A mutant suggested the formation of a hole in the channel. In the case of A602L and V605L mutants, the efficiency of ammonia transfer decreased to 50% of the value of the native protein. None of the mutants were, however, able to use exogenous ammonia as a substrate.
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FEBS Letters, Wiley, 2007, 581 (16), pp.2981-2987
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https://hal.inria.fr/inria-00188598
Contributeur : Malika Smail-Tabbone <>
Soumis le : dimanche 18 novembre 2007 - 22:09:19
Dernière modification le : mercredi 25 juillet 2018 - 10:52:02

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  • HAL Id : inria-00188598, version 1

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Nicolas Floquet, Stéphane Mouilleron, Rasha Daher, Bernard Maigret, Bernard Badet, et al.. Ammonia channeling in bacterial glucosamine-6-phosphate synthase (Glms): molecular dynamics simulations and kinetic studies of protein mutants. FEBS Letters, Wiley, 2007, 581 (16), pp.2981-2987. 〈inria-00188598〉

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