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Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin

VanA-type resistance to glycopeptide antibiotics in clinical enterococci is regulated by the VanS(A)R(A) two-component signal transduction system. The nature of the molecular ligand that is recognised by the VanS(A) sensory component has not hitherto been identified. Here we employ purified, intact...

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Detalles Bibliográficos
Autores principales: Phillips-Jones, Mary K., Channell, Guy, Kelsall, Claire J., Hughes, Charlotte S., Ashcroft, Alison E., Patching, Simon G., Dinu, Vlad, Gillis, Richard B., Adams, Gary G., Harding, Stephen E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5387412/
https://www.ncbi.nlm.nih.gov/pubmed/28397853
http://dx.doi.org/10.1038/srep46180
Descripción
Sumario:VanA-type resistance to glycopeptide antibiotics in clinical enterococci is regulated by the VanS(A)R(A) two-component signal transduction system. The nature of the molecular ligand that is recognised by the VanS(A) sensory component has not hitherto been identified. Here we employ purified, intact and active VanS(A) membrane protein (henceforth referred to as VanS) in analytical ultracentrifugation experiments to study VanS oligomeric state and conformation in the absence and presence of vancomycin. A combination of sedimentation velocity and sedimentation equilibrium in the analytical ultracentrifuge (SEDFIT, SEDFIT-MSTAR and MULTISIG analysis) showed that VanS in the absence of the ligand is almost entirely monomeric (molar mass M = 45.7 kDa) in dilute aqueous solution with a trace amount of high molar mass material (M ~ 200 kDa). The sedimentation coefficient s suggests the monomer adopts an extended conformation in aqueous solution with an equivalent aspect ratio of ~(12 ± 2). In the presence of vancomycin over a 33% increase in the sedimentation coefficient is observed with the appearance of additional higher s components, demonstrating an interaction, an observation consistent with our circular dichroism measurements. The two possible causes of this increase in s – either a ligand induced dimerization and/or compaction of the monomer are considered.