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 VanSARA two-component signal transduction system. The nature of the molecular ligand that is recognised by the VanSA sensory component has not hitherto been identified. Here we employ purified, intact and ac...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
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Nature Publishing Group
2017
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| Online Access: | https://eprints.nottingham.ac.uk/43665/ |
| _version_ | 1848796739480322048 |
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| author | Phillips-Jones, Mary K. Channell, Guy Andrew Kelsall, Claire J. Hughes, Charlotte S. Ashcroft, Alison E. Patching, Simon G. Dinu, Vlad Gillis, Richard B. Adams, Gary G. Harding, Stephen E. |
| author_facet | Phillips-Jones, Mary K. Channell, Guy Andrew Kelsall, Claire J. Hughes, Charlotte S. Ashcroft, Alison E. Patching, Simon G. Dinu, Vlad Gillis, Richard B. Adams, Gary G. Harding, Stephen E. |
| author_sort | Phillips-Jones, Mary K. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | VanA-type resistance to glycopeptide antibiotics in clinical enterococci is regulated by the VanSARA two-component signal transduction system. The nature of the molecular ligand that is recognised by the VanSA sensory component has not hitherto been identified. Here we employ purified, intact and active VanSA 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. |
| first_indexed | 2025-11-14T19:52:46Z |
| format | Article |
| id | nottingham-43665 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:52:46Z |
| publishDate | 2017 |
| publisher | Nature Publishing Group |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-436652020-05-04T18:41:43Z https://eprints.nottingham.ac.uk/43665/ Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin Phillips-Jones, Mary K. Channell, Guy Andrew Kelsall, Claire J. Hughes, Charlotte S. Ashcroft, Alison E. Patching, Simon G. Dinu, Vlad Gillis, Richard B. Adams, Gary G. Harding, Stephen E. VanA-type resistance to glycopeptide antibiotics in clinical enterococci is regulated by the VanSARA two-component signal transduction system. The nature of the molecular ligand that is recognised by the VanSA sensory component has not hitherto been identified. Here we employ purified, intact and active VanSA 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. Nature Publishing Group 2017-04-11 Article PeerReviewed Phillips-Jones, Mary K., Channell, Guy Andrew, Kelsall, Claire J., Hughes, Charlotte S., Ashcroft, Alison E., Patching, Simon G., Dinu, Vlad, Gillis, Richard B., Adams, Gary G. and Harding, Stephen E. (2017) Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin. Scientific Reports, 7 . 46180/1-46180/12. ISSN 2045-2322 https://doi.org/10.1038/srep46180 doi:10.1038/srep46180 doi:10.1038/srep46180 |
| spellingShingle | Phillips-Jones, Mary K. Channell, Guy Andrew Kelsall, Claire J. Hughes, Charlotte S. Ashcroft, Alison E. Patching, Simon G. Dinu, Vlad Gillis, Richard B. Adams, Gary G. Harding, Stephen E. Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin |
| title | Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin |
| title_full | Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin |
| title_fullStr | Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin |
| title_full_unstemmed | Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin |
| title_short | Hydrodynamics of the VanA-type VanS histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin |
| title_sort | hydrodynamics of the vana-type vans histidine kinase: an extended solution conformation and first evidence for interactions with vancomycin |
| url | https://eprints.nottingham.ac.uk/43665/ https://eprints.nottingham.ac.uk/43665/ https://eprints.nottingham.ac.uk/43665/ |