h v 1: how a voltage-sensor may form a channel younes mokrab biophysical society 53 rd annual...
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Hv1: How A Voltage-sensor May Form A
ChannelYounes Mokrab
Biophysical Society 53rd Annual Meeting
2 Mar 2009
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Hv1: a VS with proton channel activity
Ramsey et al. 2006Schilling et al. 2002
H+
S1 S2 S3 S4
+++
• Exact function not known but enriched in immune tissue.
• Topology similar to VS domains of voltage-gated ion channels.
• In response to depolarising potential, it conducts an outward H+ current, with similar characteristics to GvH+.
• Conduction pore and molecular mechanism is still unknown
Istep
-Itail
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Homology open-state models for Hv1 and simulation of insertion
into bilayer
Bond et al. 2006, Jiang et al. 2002, Jiang et al. 2003, Long et al. 2007
4:1
Full-atom Coarse-grain
Protein
Lipid
Water
Ion
Hv1 homology models based on x-ray structures of KvAP, Kv1.2 and Kv1.2-2.1 chimera.
Self assembly in POPC for 320 ns
Protein inserted to membrane
Coarse-grain (CG) models
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Full-atom simulations based on CG configuration of protein in bilayer
Per-residue fluctuationBackbone RMSD versus time
Conversion of system to full-atom using LSF
Full-atom simulation for 20 ns
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Hydrophilic residues cluster along the centre of the Hv1 bundle
Chimera-based Kv1.2-based kvAP-based
• Models might capture different open-state conformations, all showing charged/polar residues which line a potential conduction pathway for H+ during Hv1 activation.
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A water-permeable pore forms along central axis of Hv1
• Analysis of water density during simulation shows water penetration of Hv1 along its central axis in the models, forming a potential water permeable pore.
Chimera-based Kv1.2-based kvAP-based
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Water molecules are coordinated by polar and charged residues
• A number of charged side chains are identified which may be involved in direct H+ translocation, and/or polar side chains in general might coordinate a ‘proton wire’.
• In silico and In vivo mutagenesis experiment in progress to test the roles of specific residues.
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Water pore absent from VS homologues
Chimera Kv1.2 kvAP
• Simulations of other voltage-gated channels show considerably-reduced water penetration along the central axis of the VS domains.
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Pore-lining residues unique to Hv1
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Conclusions and future directions
• Combined CG-AT simulations of Hv1 open-state models in solvated lipid bilayer reveals a central water-permeable pore which seems to be absent from other VS proteins.
• High water density sites in a potential ‘gating pore’ in Hv1 appear to be coordinated by uniquely conserved polar and charged residues.
• Role of specific residues in water coordination is being tested by in silico and in vivo site-directed mutagenesis.
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Acknowledgments
• Professor Mark Sansom, SBCB, University of Oxford.
• Dr Scott Ramsey and Professor David Clapham, Harvard Medical School (Poster 3415-Pos B462, Wed 4 Mar 1-3 pm).
• Dr Zara Sands, AstraZeneca, Sweden.
• Dr Kathryn Scott, Dr Phil Stansfeld, Dr Kaihsu Tai and members of SBCB.