Strict coupling between CFTR's catalytic cycle and gating of its Cl - ion pore revealed by distributions of open channel burst durations

László Csanády, Paola Vergani, David C. Gadsby

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

CFTR, the ABC protein defective in cystic fibrosis, functions as an anion channel. Once phosphorylated by protein kinase A, a CFTR channel is opened and closed by events at its two cytosolic nucleotide binding domains (NBDs). Formation of a head-to-tail NBD1/NBD2 heterodimer, by ATP binding in two interfacial composite sites between conservedWalker A and B motifs of one NBD and the ABC-specific signature sequence of the other, has been proposed to trigger channel opening. ATP hydrolysis at the only catalytically competent interfacial site is suggested to then destabilize the NBD dimer and prompt channel closure. But this gating mechanism, and how tightly CFTR channel opening and closing are coupled to its catalytic cycle, remains controversial. Here we determine the distributions of open burst durations of individual CFTR channels, and use maximum likelihood to evaluate fits to equilibrium and nonequilibrium mechanisms and estimate the rate constants that govern channel closure. We examine partially and fully phosphorylated wild-type CFTR channels, and two mutant CFTR channels, each bearing a deleterious mutation in one or other composite ATP binding site. We show that the wild-type CFTR channel gating cycle is essentially irreversible and tightly coupled to the ATPase cycle, and that this coupling is completely destroyed by the NBD2Walker B mutation D1370N but only partially disrupted by the NBD1 Walker A mutation K464A.

Original languageEnglish
Pages (from-to)1241-1246
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number3
DOIs
Publication statusPublished - Jan 19 2010

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Keywords

  • ATPase cycle
  • Maximum likelihood
  • Nonequilibrium
  • Phosphorylation
  • Walker motifs

ASJC Scopus subject areas

  • General

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