Department of Anesthesiology

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Synergistic substrate binding determines the stoichiometry of transport of a prokaryotic H(+)/Cl(-) exchanger.

TitleSynergistic substrate binding determines the stoichiometry of transport of a prokaryotic H(+)/Cl(-) exchanger.
Publication TypeJournal Article
Year of Publication2012
AuthorsPicollo A, Xu Y, Johner N, Bernèche S, Accardi A
JournalNat Struct Mol Biol
Volume19
Issue5
Pagination525-31, S1
Date Published2012 May
ISSN1545-9985
KeywordsChloride Channels, Chlorides, Chlorine, Escherichia coli, Escherichia coli Proteins, Models, Molecular, Mutation, Protein Binding, Protons, Substrate Specificity, Thermodynamics
Abstract

Active exchangers dissipate the gradient of one substrate to accumulate nutrients, export xenobiotics and maintain cellular homeostasis. Mechanistic studies have suggested that two fundamental properties are shared by all exchangers: substrate binding is antagonistic, and coupling is maintained by preventing shuttling of the empty transporter. The CLC H(+)/Cl(-) exchangers control the homeostasis of cellular compartments in most living organisms, but their transport mechanism remains unclear. We show that substrate binding to CLC-ec1 is synergistic rather than antagonistic: chloride binding induces protonation of a crucial glutamate. The simultaneous binding of H(+) and Cl(-) gives rise to a fully loaded state that is incompatible with conventional transport mechanisms. Mutations in the Cl(-) transport pathway identically alter the stoichiometries of H(+)/Cl(-) exchange and binding. We propose that the thermodynamics of synergistic substrate binding, rather than the kinetics of conformational changes and ion binding, determine the stoichiometry of transport.

DOI10.1038/nsmb.2277
Alternate JournalNat. Struct. Mol. Biol.
PubMed ID22484316
PubMed Central IDPMC3348462
Grant List1R01GM085232 / GM / NIGMS NIH HHS / United States
R01 GM085232 / GM / NIGMS NIH HHS / United States
R01 GM085232-01A1 / GM / NIGMS NIH HHS / United States