Our long-term goal is to understand the physiological and pathophysiological role of the BK-type voltage and Ca2+-activated K+ channels. BK channels modulate physiological processes that involve Ca2+ signaling in many tissues, such as muscle contraction, renal function and neural transmission. These channels are composed of the pore-forming, voltage- and Ca2+ -sensing a subunit that is encoded by a single Slo1 gene and four types of auxiliary ? subunit (?1-4). Each of these ? subunits modulates functional properties of the Slo1 channel with distinct characteristics and tissue-specific expression. Thus, ? subunits help to define the phenotypes and physiological roles of BK channels in various tissues. To achieve our long-term goal, it is necessary to study the molecular mechanism of the interaction between Slo1 and the ? subunits. Previous publications and our preliminary data demonstrate that, although all four types of ? subunit share similar structural features, the mechanisms of their modulation of Slo1 channels differ in two key aspects: 1) they target different molecular components and processes in Slo1 that are important in channel gating, and 2) they have different amino acids or motifs (active sites) that are critical for altering gating of Slo1 channels. Based on these preliminary results, we will use the methods of electrophysiology, mutagenesis, chemical modification and kinetic modeling to achieve the following specific aims: I. To identify the molecular targets of ? subunits modulation in Slo1 channels. II. To identify the active sites of ? subunits. III. To examine if ? subunits affect the interaction between the membrane-spanning and the cytosolic domains that is critical for BK channel gating.
Aim III will establish a link between the ? subunit association, as studied in Aims I and II, with a structural mechanism of BK channel gating that has been revealed recently. This study will identify amino acids and structural motifs important for BK channel gating and reveal the nature of the interactions between Slo1 and ? subunits. It will lay the foundation for understanding the molecular basis of BK channel related pathological conditions, such as epilepsy and hypertension, and provide the target and rationale for their treatment. ? subunits of K+ channels with transmembrane segment, such as the KCNE family and BK ? subunits modulate the gating properties of their respective a subunits, which are key to the physiological roles of these channels. Our preliminary studies suggest that some of these ? subunits may affect channel gating through common mechanisms. Therefore, this study will provide insights to these common mechanisms of K+ channel function.

Public Health Relevance

This study will identify amino acids and structural motifs important for BK channel gating and reveal the nature of the interactions between Slo1 and ? subunits. It will lay the foundation for understanding the molecular basis of BK channel related pathological conditions, such as epilepsy and hypertension, and provide the target and rationale for their treatment.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS060706-04
Application #
8410089
Study Section
Biophysics of Neural Systems Study Section (BPNS)
Program Officer
Silberberg, Shai D
Project Start
2010-02-15
Project End
2014-01-31
Budget Start
2013-02-01
Budget End
2014-01-31
Support Year
4
Fiscal Year
2013
Total Cost
$314,445
Indirect Cost
$107,573
Name
Washington University
Department
Biomedical Engineering
Type
Schools of Engineering
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Kasimova, Marina A; Zaydman, Mark A; Cui, Jianmin et al. (2015) PIPâ‚‚-dependent coupling is prominent in Kv7.1 due to weakened interactions between S4-S5 and S6. Sci Rep 5:7474
Varga, Zoltan; Zhu, Wandi; Schubert, Angela R et al. (2015) Direct Measurement of Cardiac Na+ Channel Conformations Reveals Molecular Pathologies of Inherited Mutations. Circ Arrhythm Electrophysiol 8:1228-39
Zaydman, Mark A; Kasimova, Marina A; McFarland, Kelli et al. (2014) Domain-domain interactions determine the gating, permeation, pharmacology, and subunit modulation of the IKs ion channel. Elife 3:e03606
Li, Min; Chang, Shan; Yang, Longjin et al. (2014) Conopeptide Vt3.1 preferentially inhibits BK potassium channels containing β4 subunits via electrostatic interactions. J Biol Chem 289:4735-42
Zhang, Guohui; Yang, Huanghe; Liang, Hongwu et al. (2014) A charged residue in S4 regulates coupling among the activation gate, voltage, and Ca2+ sensors in BK channels. J Neurosci 34:12280-8
Zaydman, Mark A; Cui, Jianmin (2014) PIP2 regulation of KCNQ channels: biophysical and molecular mechanisms for lipid modulation of voltage-dependent gating. Front Physiol 5:195
Zaydman, Mark A; Silva, Jonathan R; Delaloye, Kelli et al. (2013) Kv7.1 ion channels require a lipid to couple voltage sensing to pore opening. Proc Natl Acad Sci U S A 110:13180-5
Li, Yang; Gao, Junyuan; Lu, Zhongju et al. (2013) Intracellular ATP binding is required to activate the slowly activating K+ channel I(Ks). Proc Natl Acad Sci U S A 110:18922-7
Lee, Hsiang-Chun; Rudy, Yoram; Po-Yuan, Phd et al. (2013) Modulation of KCNQ1 alternative splicing regulates cardiac IKs and action potential repolarization. Heart Rhythm 10:1220-8
Yang, Junqiu; Yang, Huanghe; Sun, Xiaohui et al. (2013) Interaction between residues in the Mg2+-binding site regulates BK channel activation. J Gen Physiol 141:217-28

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