Article Dans Une Revue Nature Communications Année : 2024

3D variability analysis reveals a hidden conformational change controlling ammonia transport in human asparagine synthetase

Résumé

Advances in X-ray crystallography and cryogenic electron microscopy (cryo-EM) offer the promise of elucidating functionally relevant conformational changes that are not easily studied by other biophysical methods. Here we show that 3D variability analysis (3DVA) of the cryo-EM map for wild-type (WT) human asparagine synthetase (ASNS) identifies a functional role for the Arg-142 side chain and test this hypothesis experimentally by characterizing the R142I variant in which Arg-142 is replaced by isoleucine. Support for Arg-142 playing a role in the intramolecular translocation of ammonia between the active site of the enzyme is provided by the glutamine-dependent synthetase activity of the R142 variant relative to WT ASNS, and MD simulations provide a possible molecular mechanism for these findings. Combining 3DVA with MD simulations is a generally applicable approach to generate testable hypotheses of how conformational changes in buried side chains might regulate function in enzymes.
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hal-04910045 , version 1 (24-02-2025)
hal-04910045 , version 2 (24-02-2025)

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Adriana Coricello, Alanya Nardone, Antonio Lupia, Carmen Gratteri, Matthijn Vos, et al.. 3D variability analysis reveals a hidden conformational change controlling ammonia transport in human asparagine synthetase. Nature Communications, 2024, 15 (1), pp.10538. ⟨10.1038/s41467-024-54912-9⟩. ⟨hal-04910045v2⟩
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