Abstract

To understand the role of the Slack gene in neurons and other excitable cells, it is essential to characterize the heteromeric channels resulting from the co-expression of Slack and Slo subunits. In this project we will identify the subunit stoichiometries that result in functional channels and characterize these channels in regard to their voltage and Ca sensitivity and conductance properties. We will also test for the functional properties of """"""""core/tail"""""""" constructs of Slack subunits created in Project 2. Further, we will establish systems for expression and purification of Slack and Slo protein; with purified, reconstituted channel protein, we will address the three-dimensional structure of the Maxi-K channel complex. Heretofore the only direct structural information about the six-transmembrane-segment family of ion channel proteins consists of crystal structures of cytoplasmic domains and low-resolution, projection EM images of negatively-stained channel proteins. We will obtain three-dimensional structural information at moderate resolution of Maxi-K channels using the combination of cryo-electronmicroscopy and novel single-particle reconstruction techniques.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Program Projects (P01)
Project #
1P01NS042202-01
Application #
6552302
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Project Start
2001-09-04
Project End
2006-08-31
Budget Start
Budget End
Support Year
1
Fiscal Year
2001
Total Cost
Indirect Cost

  Institution

Name
Yale University
Department
Type
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520

  Publications

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

Showing the most recent 10 out of 20 publications