Functional study of cytoplasmic loops of human skeletal muscle chloride channel, hClC-1

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dc.contributor.authorMa, L.
dc.contributor.authorRychkov, G.
dc.contributor.authorBretag, A.
dc.date.issued2009
dc.description.abstractThe membrane-resident domain of chloride channels and transporters of the CLC family is composed of 18 alpha-helices (designated A to R) connected sequentially by extracellular and intracellular loops, whose functional characteristics are generally unclear. To study the relevance of the intracellular loops linking helices D and E, F and G, H and I and J and K, alanine-exchange mutagenesis, split channel strategy, GST (glutathione transferase)-pull-down methods and whole-cell patch-clamp recordings were used. We investigated the possible roles of these loops in binding to the cytoplasmic, carboxyl tail (C-tail) of the protein, as well as their physiological roles in channel function. Although other interacting positions are conceivable, our results indicate that there is unlikely to be significant binding between the C-tail and any one of these individual cytoplasmic loops. Particular loops might, however, be essential for some channel characteristics. For example, alanine-exchange mutation of the loop linking helix D to E eliminated channel currents; of the loop linking helix H to I caused a significant shift of the open probability of fast gating (P(o)(f)), towards more positive voltages; and of the loop linking helix J to K locked the common gating of hClC-1 open. Therefore, the gating mechanisms of hClC-1 might not only involve the helices and the C-tail, but also involve some of the loops.
dc.description.statementofresponsibilityLinlin Ma, Grigori Y. Rychkov and Allan H. Bretag
dc.description.urihttp://www.elsevier.com/wps/find/journaldescription.cws_home/395/description#description
dc.identifier.citationThe International Journal of Biochemistry and Cell Biology, 2009; 41(6):1402-1409
dc.identifier.doi10.1016/j.biocel.2008.12.006
dc.identifier.issn1357-2725
dc.identifier.issn1878-5875
dc.identifier.orcidRychkov, G. [0000-0002-2788-2977]
dc.identifier.urihttp://hdl.handle.net/2440/51484
dc.language.isoen
dc.publisherPergamon-Elsevier Science Ltd
dc.source.urihttps://doi.org/10.1016/j.biocel.2008.12.006
dc.subjectMuscle, Skeletal
dc.subjectCells, Cultured
dc.subjectCytoplasm
dc.subjectHumans
dc.subjectAlanine
dc.subjectChloride Channels
dc.subjectBlotting, Western
dc.subjectTransfection
dc.subjectGene Expression
dc.subjectProtein Structure, Tertiary
dc.subjectStructure-Activity Relationship
dc.titleFunctional study of cytoplasmic loops of human skeletal muscle chloride channel, hClC-1
dc.typeJournal article
pubs.publication-statusPublished

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