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Movement of hCIC-1 C-termini during common gating and limits on their cytoplasmic location

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dc.contributor.authorMa, L.
dc.contributor.authorRychkov, G.
dc.contributor.authorBykova, E.
dc.contributor.authorZheng, J.
dc.contributor.authorBretag, A.
dc.date.issued2011
dc.description.abstractFunctionally, the dimeric human skeletal muscle chloride channel hClC-1 is characterized by two distinctive gating processes, fast (protopore) gating and slow (common) gating. Of these, common gating is poorly understood, but extensive conformational rearrangement is suspected. To examine this possibility, we used FRET (fluorescence resonance energy transfer) and assessed the effects of manipulating the common-gating process. Closure of the common gate was accompanied by a separation of the C-termini, whereas, with opening, the C-termini approached each other more closely. These movements were considerably smaller than those seen in ClC-0. To estimate the C-terminus depth within the cytoplasm we constructed a pair of split hClC-1 fragments tagged extracellularly and intracellularly respectively. These not only combined appropriately to rescue channel function, but we detected positive FRET between them. This restricts the C-termini of hClC-1 to a position close to its membrane-resident domain. From mutants in which fast or common gating were affected, FRET revealed a close linkage between the two gating processes with the carboxyl group of Glu²³² apparently acting as the final effector for both.
dc.description.statementofresponsibilityLinlin Ma, Grigori Y. Rychkov, Katerina A. Bykova, Jie Zheng and Allan H. Bretag
dc.identifier.citationBiochemical Journal, 2011; 436(2):415-428
dc.identifier.doi10.1042/BJ20102153
dc.identifier.issn0264-6021
dc.identifier.issn1470-8728
dc.identifier.orcidRychkov, G. [0000-0002-2788-2977]
dc.identifier.urihttp://hdl.handle.net/2440/68920
dc.language.isoen
dc.publisherPortland Press
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/453580
dc.rights© The Authors Journal compilation © 2011 Biochemical Society
dc.source.urihttps://doi.org/10.1042/BJ20102153
dc.subjectC-terminus
dc.subjectcommon gating
dc.subjectconformation change
dc.subjectcytoplasmic domain
dc.subjecthuman skeletal muscle chloride channel (hClC-1)
dc.subjectfluorescence resonance energy transfer (FRET).
dc.titleMovement of hCIC-1 C-termini during common gating and limits on their cytoplasmic location
dc.typeJournal article
pubs.publication-statusPublished

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