Mycobacterium tuberculosis NmtR harbors a Nickel sensing site with parallels to Escherichia coli RcnR

Hermes Reyes-Caballero, Chul Won Lee, David P. Giedroc

Research output: Contribution to journalArticle

Abstract

Mycobacterium tuberculosis NmtR is a Ni(II)/Co(II)-sensing metalloregulatory protein from the extensively studied ArsR/SmtB family. Two Ni(II) ions bind to the NmtR dimer to form octahedral coordination complexes with the following stepwise binding affinities: K Ni1 = (1.2 ± 0.1) ×10 10 M -1, and K Ni2 = (0.7 ± 0.4) ×10 10 M -1 (pH 7.0). A glutamine scanning mutagenesis approach reveals that Asp91, His93, His104, and His107, all contained within the C-terminal α5 helix, and His3 as part of the conserved α-NH 2-Gly2-His3-Gly4 motif at the N-terminus make significant contributions to the magnitude of K Ni. In contrast, substitution of residues from the C-terminal region, His109, Asp114, and His116, previously implicated in Ni(II) binding and metalloregulation in cells, gives rise to wild-type K Ni and Ni(II)-dependent allosteric coupling free energies. Interestingly, deletion of residues 112-120 from the C-terminal region (Δ111 NmtR) reduces the Ni(II) binding stoichiometry to one per dimer and greatly reduces Ni(II) responsiveness. H3Q and Δ111 NmtRs also show clear perturbations in the rank order of metal responsiveness to Ni(II), Co(II), and Zn(II) that is distinct from that of wild-type NmtR. 15N relaxation experiments with apo-NmtR reveal that both N-terminal (residues 2-14) and C- terminal (residues 110-120) regions are unstructured in solution, and this property likely dictates the metal specificity profile characteristic of the Ni(II) sensor NmtR relative to other ArsR family regulators.

Original languageEnglish (US)
Pages (from-to)7941-7952
Number of pages12
JournalBiochemistry
Volume50
Issue number37
DOIs
StatePublished - Sep 20 2011
Externally publishedYes

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Ports and harbors
Nickel
Mycobacterium tuberculosis
Dimers
Escherichia coli
Metals
Mutagenesis
Coordination Complexes
Glutamine
Stoichiometry
Free energy
Substitution reactions
Ions
Scanning
Sensors
Proteins
Experiments

ASJC Scopus subject areas

  • Biochemistry

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Mycobacterium tuberculosis NmtR harbors a Nickel sensing site with parallels to Escherichia coli RcnR. / Reyes-Caballero, Hermes; Lee, Chul Won; Giedroc, David P.

In: Biochemistry, Vol. 50, No. 37, 20.09.2011, p. 7941-7952.

Research output: Contribution to journalArticle

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