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- W2158340750 abstract "Ca2+ release-activated Ca2+ (CRAC) channels mediate Ca2+ entry in response to store depletion in a variety of cell types. Endogenous CRAC channels in mammals are formed by a homomeric assembly of Orai1 proteins. Earlier functional studies involving chemical cross-linking of different numbers of wild-type Orai1 subunits, electrophysiological recordings and single molecule fluorescence analysis techniques as well as high-resolution electron microscopic examination of purified Orai proteins strongly indicated that functional Orai channels were most likely tetramers. In contrast, study of crystals containing holo-dOrai channel complexes from Drosophila melanogaster, revealed that a single channel complex contains six dOrai subunits. The hexameric CRAC channel stoichiometry was further supported by cross-linking and size-exclusion chromatography studies of the Drosophila Orai. Recent functional study revealed that expression of concatenated hexameric Orai1 channel produces a non-selective cation channel with biophysical properties essentially different from those of endogenous CRAC channels or channels formed by expressed concatenated tetrameric Orai1 proteins. Thus, the studies reported are far from conclusive but they do indicate the need for further investigation of Orai1 channel stoichiometry. Accordingly, we generated structural models of Orai1 ion conduction pathway formed by tetrameric or hexameric assembly of Orai1 pore-lining TM1 segments using Rosetta fold and dock protocol. Based on available experimental data we constrained proximity of several key residues lining Orai1 ion conduction pathway during simulations. Among the lowest energy and most frequently sampled conformations of Orai TM1 hexamers generated by Rosetta, we identified structural models that were in close agreement with Orai structure in the transmembrane region proposed based on crystallographic data analysis. Our preliminary models of Orai TM1 tetramers suggest alternative structural topology forming ion conduction pathway, which may account for the differences in ionic selectivity of hexameric and tetrameric Orai1 channels." @default.
- W2158340750 created "2016-06-24" @default.
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- W2158340750 date "2014-01-01" @default.
- W2158340750 modified "2023-09-29" @default.
- W2158340750 title "Structural Modeling of Hexameric and Tetrameric Ion Conduction Pathways of Orai1 Channel" @default.
- W2158340750 doi "https://doi.org/10.1016/j.bpj.2013.11.1816" @default.
- W2158340750 hasPublicationYear "2014" @default.
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