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• W1970596908 startingPage "3079" @default.
• W1970596908 abstract "Activation of BKCa channels by direct Ca2+ binding and membrane depolarization occur via independent and additive molecular processes. The “calcium bowl” domain is critically involved in Ca2+-dependent gating, and we have hypothesized that a sequence within this domain may resemble an EF hand motif. Using a homology modeling strategy, it was observed that a single Ca2+ ion may be coordinated by the oxygen-containing side chains of residues within the calcium bowl (i.e., 912ELVNDTNVQFLD923). To examine these predictions directly, alanine-substituted BKCa channel mutants were expressed in HEK 293 cells and the voltage and Ca2+ dependence of macroscopic currents were examined in inside-out membrane patches. Over the range of 1–10 μM free Ca2+, single point mutations (i.e., E912A and D923A) produced rightward shifts in the steady-state conductance-voltage relations, whereas the mutants N918A or Q920A had no effect on Ca2+-dependent gating. The double mutant E912A/D923A displayed a synergistic shift in Ca2+-sensitive gating, as well as altered kinetics of current activation/deactivation. In the presence of 1, 10, and 80 mM cytosolic Mg2+, this double mutation significantly reduced the Ca2+-induced free energy change associated with channel activation. Finally, mutations that altered sensitivity of the holo-channel to Ca2+ also reduced direct 45Ca binding to the calcium bowl domain expressed as a bacterial fusion protein. These findings, along with other recent data, are considered in the context of the calcium bowl’s high affinity Ca2+ sensor and the known properties of EF hands." @default.
• W1970596908 created "2016-06-24" @default.
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• W1970596908 date "2005-11-01" @default.
• W1970596908 modified "2023-09-25" @default.
• W1970596908 title "Homology Modeling Identifies C-Terminal Residues that Contribute to the Ca2+ Sensitivity of a BKCa Channel" @default.
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• W1970596908 doi "https://doi.org/10.1529/biophysj.105.063610" @default.
• W1970596908 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/1366805" @default.
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