ArticleProceedings of the National Academy of Sciences of the United States of America2001
Ca2+-binding activity of a COOH-terminal fragment of the Drosophila BK channel involved in Ca2+-dependent activation.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 57 papers.
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57 citing papers in PubMed, 92 citations in OpenAlex.
- Molecular determinants of cholesterol binding to BK channel-forming slo1 subunits.Archives of biochemistry and biophysics · 2026Article
- A Calcium Sensor Discovered in Bluetongue Virus Nonstructural Protein 2 Is Critical for Virus Replication.Journal of virology · 2020Article
- Threading the biophysics of mammalian Slo1 channels onto structures of an invertebrate Slo1 channel.The Journal of general physiology · 2017Review
- Interactions of divalent cations with calcium binding sites of BK channels reveal independent motions within the gating ring.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Two helices in the third intracellular loop determine anoctamin 1 (TMEM16A) activation by calcium.Pflugers Archiv : European journal of physiology · 2015Article
- Putative calcium-binding domains of the Caenorhabditis elegans BK channel are dispensable for intoxication and ethanol activation.Genes, brain, and behavior · 2015Article
- BK channels: multiple sensors, one activation gate.Frontiers in physiology · 2015Review
- BK channel activation by tungstate requires the β1 subunit extracellular loop residues essential to modulate voltage sensor function and channel gating.Pflugers Archiv : European journal of physiology · 2014Article
- Interaction of the BKCa channel gating ring with dendrotoxins.Channels (Austin, Tex.) · 2014Article
- Transduction of voltage and Ca2+ signals by Slo1 BK channels.Physiology (Bethesda, Md.) · 2013Review
- State-dependent FRET reports calcium- and voltage-dependent gating-ring motions in BK channels.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
- Relative transmembrane segment rearrangements during BK channel activation resolved by structurally assigned fluorophore-quencher pairing.The Journal of general physiology · 2012Article
- Barium ions selectively activate BK channels via the Ca2+-bowl site.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- The contribution of RCK domains to human BK channel allosteric activation.The Journal of biological chemistry · 2012Article
- Metal-driven operation of the human large-conductance voltage- and Ca2+-dependent potassium channel (BK) gating ring apparatus.The Journal of biological chemistry · 2011Article
- Cysteine residue 911 in C-terminal tail of human BK(Ca)α channel subunit is crucial for its activation by carbon monoxide.Pflugers Archiv : European journal of physiology · 2011Article
- BK-type calcium-activated potassium channels: coupling of metal ions and voltage sensing.The Journal of physiology · 2010Review
- Relative motion of transmembrane segments S0 and S4 during voltage sensor activation in the human BK(Ca) channel.The Journal of general physiology · 2010Article
- The RCK1 domain of the human BKCa channel transduces Ca2+ binding into structural rearrangements.The Journal of general physiology · 2010Article
- Structure of the human BK channel Ca2+-activation apparatus at 3.0 A resolution.Science (New York, N.Y.) · 2010Article
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3 authors at 1 institution in 1 country.
Funding
Abstract
Mutational and biophysical analysis suggests that an intracellular COOH-terminal domain of the large conductance Ca(2+)-activated K(+) channel (BK channel) contains Ca(2+)-binding site(s) that are allosterically coupled to channel opening. However the structural basis of Ca(2+) binding to BK channels is unknown. To pursue this question, we overexpressed the COOH-terminal 280 residues of the Drosophila slowpoke BK channel (Dslo-C280) as a FLAG- and His(6)-tagged protein in Escherichia coli. We purified Dslo-C280 in soluble form and used a (45)Ca(2+)-overlay protein blot assay to detect Ca(2+) binding. Dslo-C280 exhibits specific binding of (45)Ca(2+) in comparison with various control proteins and known EF-hand Ca(2+)-binding proteins. A mutation (D5N5) of Dslo-C280, in which five consecutive Asp residues of the "Ca-bowl" motif are changed to Asn, reduces (45)Ca(2+)-binding activity by 56%. By electrophysiological assay, the corresponding D5N5 mutant of the Drosophila BK channel expressed in HEK293 cells exhibits lower Ca(2+) sensitivity for activation and a shift of approximately +80 mV in the midpoint voltage for activation. This effect is associated with a decrease in the Hill coefficient (N) for activation by Ca(2+) and a reduction in apparent Ca(2+) affinity, suggesting the loss of one Ca(2+)-binding site per monomer. These results demonstrate a functional correlation between Ca(2+) binding to a specific region of the BK protein and Ca(2+)-dependent activation, thus providing a biochemical approach to study this process.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.