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Discovering Targetable Conformation of RhoA Mutant by Integrating Native Mass Spectrometry, Ultraviolet Photodissociation, and X-ray Diffraction - PubMed

4 hours ago
  • #RhoA mutant
  • #drug discovery
  • #conformational dynamics
  • Pathogenic mutations in Ras superfamily proteins present challenges for drug development due to subtle, dynamic conformational changes.
  • Integration of X-ray crystallography, native mass spectrometry (nMS), and ultraviolet photodissociation (UVPD) revealed a cryptic conformation in the RhoA Y42C mutant.
  • nMS-UVPD identified the dominant conformation, an enhanced Mg2+-locked state, overcoming ambiguities from crystallography alone.
  • The mutation impairs GTP hydrolysis, revealing a hidden druggable pocket near Cys42.
  • A covalent inhibitor was identified, targeting the newly exposed pocket.
  • This integrated approach provides a strategy for targeting dynamic, 'undruggable' protein mutants.