Thermodynamic Studies of the Mechanism of Metal Binding to the Escherichia coli Zinc Transporter YiiP

Yang Chao, Dax Fu

Research output: Contribution to journalArticle

Abstract

Sequence homology of the Escherichia coli YiiP places it within the family of cation diffusion facilitators, a family of membrane transporters that play a central role in regulating cellular zinc homeostasis. Here we describe the first thermodynamic and mechanistic studies of metal binding to a cation diffusion facilitator. Isothermal titration calorimetric analyses of the purified YiiP and binding competitions among Zn2+, Cd2+, and Hg2+ revealed a mutually competitive binding site common to three metal ions and a set of noncompetitive binding sites, including one Cd2+ site, one Hg2+ site, and at least one Zn 2+ site, to which the binding of Zn2+ exhibited partial inhibitions of both Cd2+ and Hg2+ bindings. Lowering the pH from 7.0 to 5.5 inhibited binding of Zn2+ and Cd2+ to the common site. Further, the enthalpy change of the Cd2+ binding to the common site was found to be related linearly to the ionization enthalpy of the pH buffer with a slope corresponding to the release of 1.28 H+ for each Cd2+ binding. These H+ effects are consistent with a coupled deprotonation process upon binding of Zn2+ and Cd 2+. Modification of histidine residues by diethyl pyrocarbonate specifically inhibited Zn2+ binding to the common binding site, indicating that the mechanism of binding-deprotonation coupling involves a histidine residue(s).

Original languageEnglish (US)
Pages (from-to)17173-17180
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number17
DOIs
StatePublished - Apr 23 2004
Externally publishedYes

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Thermodynamics
Escherichia coli
Deprotonation
Metals
Binding Sites
Histidine
Cations
Enthalpy
Diethyl Pyrocarbonate
Competitive Binding
Membrane Transport Proteins
Sequence Homology
Titration
Ionization
Metal ions
Zinc
Buffers
Homeostasis
Ions
zinc-binding protein

ASJC Scopus subject areas

  • Biochemistry

Cite this

Thermodynamic Studies of the Mechanism of Metal Binding to the Escherichia coli Zinc Transporter YiiP. / Chao, Yang; Fu, Dax.

In: Journal of Biological Chemistry, Vol. 279, No. 17, 23.04.2004, p. 17173-17180.

Research output: Contribution to journalArticle

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