Shaker, a model for the subfamily of voltage-dependent potassium (Kv) channels, catalyzes the transmembranous flow of potassium ions upon membrane depolarization. Kv channels are involved in a myriad of physiological processes including cardiac cell repolarization and frequency-encoding in neurons. Although this integral membrane protein has been subject to extensive biophysical characterization, details concerning the channel's activation mechanism are scant. In order to elucidate the manner in which the transmembrane electric field affects voltage sensor movement, a combination of optical and electrophysiological techniques will be utilized. The use of site-directed mutagenesis and thiol-specific fluorophores, allows specific structural modifications to be assayed via fluorescence techniques and correlated to gating charge displacement. Therefore, this research proposal will focus on two aspects of Shaker channel voltage-sensor dynamics: i.) determining equilibrium positions of S4 residues with the use of Lanthanide- Based Fluorescence Resonance Energy Transfer (LRET), and ii.) characterizing the kinetics of local conformational changes with the use of site-specific fluorescence The insight gained from this study will augment the contemporary view of ion channel biophysics and may result in a better understanding of normal physiology, in addition to the pathophysiology of numerous channelopathies.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31GM020596-03
Application #
6525706
Study Section
Special Emphasis Panel (ZRG1-IFCN-5 (02))
Program Officer
Toliver, Adolphus
Project Start
2002-08-01
Project End
Budget Start
2002-08-01
Budget End
2003-07-31
Support Year
3
Fiscal Year
2002
Total Cost
$24,749
Indirect Cost
Name
University of California Los Angeles
Department
Physiology
Type
Schools of Medicine
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Chanda, Baron; Asamoah, Osei Kwame; Bezanilla, Francisco (2004) Coupling interactions between voltage sensors of the sodium channel as revealed by site-specific measurements. J Gen Physiol 123:217-30
Asamoah, Osei Kwame; Wuskell, Joseph P; Loew, Leslie M et al. (2003) A fluorometric approach to local electric field measurements in a voltage-gated ion channel. Neuron 37:85-97