A class of calcium-activated potassium channels termed BK(Ca) channels are enriched at presynaptic neurotransmitter release sites within the brain, and are believed to control both the amount and timing of neurotransmitter release. BK(Ca) channels are encoded by the Slo gene. Recent evidence suggests that a related gene, Slack, also contributes to BK(Ca)-like channels in neurons, and that some BK(Ca) channels may be comprised of heteromultimers of Slo and Slack channels subunits. The experiments in this proposal will characterize the molecular, biophysical and structural properties of BK(Ca) channels in both the somata and presynaptic terminals of native neurons, and in transfected cells. The role of the Slo and Slack channel subunits in normal synaptic transmission, and the mechanisms of their response to hypoxia, will be determined by patch clamp measurements coupled with genetic knockout approaches. The structure and function of the large carboxy-terminal regulatory region of the Slo channel subunit with its calcium-binding sites, and its interaction with other proteins, including the Slack subunit, will be determined by biophysical and biochemical measurements and by X-ray crystallographic methods. Finally the structure of intact BK(Ca) channels will be determined by cryo-electronmicroscopy. Knowledge of the molecular properties and regulation of BK(Ca) channels in normal and hypoxic neurons, together with structural determinations of their interactions with both drugs and natural ligands, will be key in the development of more effective treatments for disorders such as epilepsy and stroke.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Program Projects (P01)
Project #
1P01NS042202-01
Application #
6369473
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Program Officer
Talley, Edmund M
Project Start
2001-09-04
Project End
2006-08-31
Budget Start
2001-09-04
Budget End
2002-08-31
Support Year
1
Fiscal Year
2001
Total Cost
$1,474,141
Indirect Cost
Name
Yale University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Takacs, Zoltan; Imredy, John P; Bingham, Jon-Paul et al. (2014) Interaction of the BKCa channel gating ring with dendrotoxins. Channels (Austin) 8:421-32
Bingham, Jon-Paul; Chun, Joycelyn B; Ruzicka, Margaret R et al. (2009) Synthesis of an iberiotoxin derivative by chemical ligation: a method for improved yields of cysteine-rich scorpion toxin peptides. Peptides 30:1049-57
Brown, Maile R; Kronengold, Jack; Gazula, Valeswara-Rao et al. (2008) Amino-termini isoforms of the Slack K+ channel, regulated by alternative promoters, differentially modulate rhythmic firing and adaptation. J Physiol 586:5161-79
Krishnan, Manoj N; Trombley, Patrick; Moczydlowski, Edward G (2008) Thermal stability of the K+ channel tetramer: cation interactions and the conserved threonine residue at the innermost site (S4) of the KcsA selectivity filter. Biochemistry 47:5354-67
Sun, Xiaolu; Yao, Hang; Zhou, Dan et al. (2008) Modulation of hSlo BK current inactivation by fatty acid esters of CoA. J Neurochem 104:1394-403
Sun, X; Zhou, D; Zhang, P et al. (2007) Beta-subunit-dependent modulation of hSlo BK current by arachidonic acid. J Neurophysiol 97:62-9
Douglas, Robert M; Miyasaka, Naoyuki; Takahashi, Kan et al. (2007) Chronic intermittent but not constant hypoxia decreases NAA/Cr ratios in neonatal mouse hippocampus and thalamus. Am J Physiol Regul Integr Comp Physiol 292:R1254-9
Yang, Bo; Desai, Rooma; Kaczmarek, Leonard K (2007) Slack and Slick K(Na) channels regulate the accuracy of timing of auditory neurons. J Neurosci 27:2617-27
Santi, Celia M; Ferreira, Gonzalo; Yang, Bo et al. (2006) Opposite regulation of Slick and Slack K+ channels by neuromodulators. J Neurosci 26:5059-68
Bingham, Jon-Paul; Bian, Shumin; Tan, Zhi-Yong et al. (2006) Synthesis of a biotin derivative of iberiotoxin: binding interactions with streptavidin and the BK Ca2+-activated K+ channel expressed in a human cell line. Bioconjug Chem 17:689-99

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