Voltage-sensing domains (VSDs) are transmembrane protein modules that detect electrical signals propagating within cell membranes. Ion channels and enzymes containing these domains play key roles in many biological processes, from the generation of the action potential in neurons and muscles, to the regulation of reactive oxygen species (ROS) during infection and inflammation. Malfunction or misexpression of VSD-containing proteins is associated with numerous diseases, such as epilepsy, periodic paralysis, cardiac arrhythmia, cancer and autoimmune disorders. Some VSDs conduct ions across the membrane under physiological conditions. Others become ion permeant under pathological conditions, as a result of mutations. The long-term goal of this study is to elucidate the mechanism underlying ion conduction through the VSD and its relationship to the general mechanism of voltage sensing. The study focuses on the voltage-gated proton channel Hv1, a protein that lacks the pore domain typical of voltage-gated sodium, potassium, and calcium channels and conducts protons through its VSD. Recent work has begun to unveil the structural organization of Hv1, but many open questions remain about the mechanisms of proton permeation, gating, and modulation of the channel. In this study we plan to answer some of these questions by using an approach that combines electrophysiological and fluorescence techniques to mutagenesis scanning and molecular dynamics simulations. Specifically, we aim at: 1) determining which parts of the VSD make up the proton pore and gate by using a novel technique of perturbation analysis recently developed in our laboratory, 2) exploring the relationship between the mechanism of proton permeation through the VSD and the mechanism of voltage sensing, using new Hv1 blockers as molecular probes, and 3) investigating the mechanisms of subunit coupling and gating modulation by accessory proteins. The proposed research will significantly expand our understanding of how VSDs sense the membrane potential, conduct ions, and interact with intracellular processes via accessory proteins. The work will also pave the way to the development of Hv1 inhibitors that can be used to address ROS overproduction typical of several cardiovascular and inflammatory disorders and will provide new insights on how mutations of VSDs lead to disease.

Public Health Relevance

Biological processes as diverse as neuronal signaling, muscle contraction, the immune response, and the heartbeat depend on the proper function of proteins containing voltage-sensing domains (VSDs). Some of these proteins conduct ions through their VSD under physiological or pathological conditions, and this project aims at determining the mechanisms underlying such ion conduction. The results of this study will help develop new drugs that reduce production of reactive oxygen species involved in chronic inflammation and cardiovascular diseases, and will provide new insights on how mutations of VSD-containing proteins lead to disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM098973-02
Application #
8290313
Study Section
Biophysics of Neural Systems Study Section (BPNS)
Program Officer
Deatherage, James F
Project Start
2011-07-01
Project End
2016-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
2
Fiscal Year
2012
Total Cost
$284,539
Indirect Cost
$94,539
Name
University of California Irvine
Department
Physiology
Type
Schools of Medicine
DUNS #
046705849
City
Irvine
State
CA
Country
United States
Zip Code
92697
Pathak, Medha M; Nourse, Jamison L; Tran, Truc et al. (2014) Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells. Proc Natl Acad Sci U S A 111:16148-53
Hong, Liang; Kim, Iris H; Tombola, Francesco (2014) Molecular determinants of Hv1 proton channel inhibition by guanidine derivatives. Proc Natl Acad Sci U S A 111:9971-6
Kim, Iris H; Hevezi, Peter; Varga, Csaba et al. (2014) Evidence for functional diversity between the voltage-gated proton channel Hv1 and its closest related protein HVRP1. PLoS One 9:e105926
Hong, Liang; Pathak, Medha M; Kim, Iris H et al. (2013) Voltage-sensing domain of voltage-gated proton channel Hv1 shares mechanism of block with pore domains. Neuron 77:274-87