K+ channels: gating mechanisms and lipid interactions
<p>Computational methods, including homology modelling, <em>in-silico</em> dockings, and molecular dynamics simulations have been used to study the functional dynamics and interactions of K<sup>+</sup> channels. Molecular models were built of the inwardly rectifying K&l...
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| Format: | Thesis |
| Language: | English |
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2013
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| _version_ | 1817933150266851328 |
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| author | Schmidt, M |
| author2 | Sansom, M |
| author_facet | Sansom, M Schmidt, M |
| author_sort | Schmidt, M |
| collection | OXFORD |
| description | <p>Computational methods, including homology modelling, <em>in-silico</em> dockings, and molecular dynamics simulations have been used to study the functional dynamics and interactions of K<sup>+</sup> channels. Molecular models were built of the inwardly rectifying K<sup>+</sup> channel Kir2.2, the bacterial homolog K<sup>+</sup> channel KirBac3.1, and the twin pore (K2P) K<sup>+</sup> channels TREK-1 and TRESK. To investigate the electrostatic energy profile of K<sup>+</sup> permeating through these homology models, continuum electrostatic calculations were performed. The primary mechanism of KirBac3.1 gating is believed to involve an opening at the helix bundle crossing (HBC). However, simulations of Kir channels have not yet revealed opening at the HBC. Here, in simulations of the new KirBac3.1-S129R X-ray crystal structure, in which the HBC was trapped open by the S129R mutation in the inner pore-lining helix (TM2), the HBC was found to exhibit considerable mobility. In a simulation of the new KirBac3.1-S129R-S205L double mutant structure, if the S129R and the S205L mutations were converted back to the wild-type serine, the HBC would close faster than in the simulations of the KirBac3.1-S129R single mutant structure. The double mutant structure KirBac3.1-S129R-S205L therefore likely represents a higher-energy state than the single mutant KirBac3.1-S129R structure, and these simulations indicate a staged pathway of gating in KirBac channels. Molecular modelling and MD simulations of the Kir2.2 channel structure demonstrated that the HBC would tend to open if the C-linker between the transmembrane and cytoplasmic domain was modelled helical. The electrostatic energy barrier for K<sup>+</sup> permeation at the helix bundle crossing was found to be sensitive to subtle structural changes in the C-linker. Charge neutralization or charge reversal of the PIP2-binding residue R186 on the C-linker decreased the electrostatic barrier for K<sup>+</sup> permeation through the HBC, suggesting an electrostatic contribution to the PIP2-dependent gating mechanism. Multi-scale simulations determined the PIP2 binding site in Kir2.2, in good agreement with crystallographic predictions. A TREK-1 homology model was built, based on the TRAAK structure. Two PIP2 binding sites were found in this TREK-1 model, at the C-terminal end, in line with existing functional data, and between transmembrane helices TM2 and TM3. The TM2-TM3 site is in reasonably good agreement with electron density attributed to an acyl tail in a recently deposited TREK-2 structure.</p> |
| first_indexed | 2024-03-07T08:18:32Z |
| format | Thesis |
| id | oxford-uuid:51dc4149-d943-4dcd-bf5b-f04130456d84 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-12-09T03:49:14Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | oxford-uuid:51dc4149-d943-4dcd-bf5b-f04130456d842024-12-08T12:28:03ZK+ channels: gating mechanisms and lipid interactionsThesishttp://purl.org/coar/resource_type/c_db06uuid:51dc4149-d943-4dcd-bf5b-f04130456d84Computational biochemistryMolecular biophysics (biochemistry)BiochemistryLife SciencesEnglishOxford University Research Archive - Valet2013Schmidt, MSansom, MTucker, S<p>Computational methods, including homology modelling, <em>in-silico</em> dockings, and molecular dynamics simulations have been used to study the functional dynamics and interactions of K<sup>+</sup> channels. Molecular models were built of the inwardly rectifying K<sup>+</sup> channel Kir2.2, the bacterial homolog K<sup>+</sup> channel KirBac3.1, and the twin pore (K2P) K<sup>+</sup> channels TREK-1 and TRESK. To investigate the electrostatic energy profile of K<sup>+</sup> permeating through these homology models, continuum electrostatic calculations were performed. The primary mechanism of KirBac3.1 gating is believed to involve an opening at the helix bundle crossing (HBC). However, simulations of Kir channels have not yet revealed opening at the HBC. Here, in simulations of the new KirBac3.1-S129R X-ray crystal structure, in which the HBC was trapped open by the S129R mutation in the inner pore-lining helix (TM2), the HBC was found to exhibit considerable mobility. In a simulation of the new KirBac3.1-S129R-S205L double mutant structure, if the S129R and the S205L mutations were converted back to the wild-type serine, the HBC would close faster than in the simulations of the KirBac3.1-S129R single mutant structure. The double mutant structure KirBac3.1-S129R-S205L therefore likely represents a higher-energy state than the single mutant KirBac3.1-S129R structure, and these simulations indicate a staged pathway of gating in KirBac channels. Molecular modelling and MD simulations of the Kir2.2 channel structure demonstrated that the HBC would tend to open if the C-linker between the transmembrane and cytoplasmic domain was modelled helical. The electrostatic energy barrier for K<sup>+</sup> permeation at the helix bundle crossing was found to be sensitive to subtle structural changes in the C-linker. Charge neutralization or charge reversal of the PIP2-binding residue R186 on the C-linker decreased the electrostatic barrier for K<sup>+</sup> permeation through the HBC, suggesting an electrostatic contribution to the PIP2-dependent gating mechanism. Multi-scale simulations determined the PIP2 binding site in Kir2.2, in good agreement with crystallographic predictions. A TREK-1 homology model was built, based on the TRAAK structure. Two PIP2 binding sites were found in this TREK-1 model, at the C-terminal end, in line with existing functional data, and between transmembrane helices TM2 and TM3. The TM2-TM3 site is in reasonably good agreement with electron density attributed to an acyl tail in a recently deposited TREK-2 structure.</p> |
| spellingShingle | Computational biochemistry Molecular biophysics (biochemistry) Biochemistry Life Sciences Schmidt, M K+ channels: gating mechanisms and lipid interactions |
| title | K+ channels: gating mechanisms and lipid interactions |
| title_full | K+ channels: gating mechanisms and lipid interactions |
| title_fullStr | K+ channels: gating mechanisms and lipid interactions |
| title_full_unstemmed | K+ channels: gating mechanisms and lipid interactions |
| title_short | K+ channels: gating mechanisms and lipid interactions |
| title_sort | k channels gating mechanisms and lipid interactions |
| topic | Computational biochemistry Molecular biophysics (biochemistry) Biochemistry Life Sciences |
| work_keys_str_mv | AT schmidtm kchannelsgatingmechanismsandlipidinteractions |