Rhizobium leguminosarum HupE is a highly-specific diffusion facilitator for nickel uptake

Marta Albareda, Agnès Rodrigue, Belén Brito, Tomás Ruiz-Argüeso, Juan Imperial, Marie Andrée Mandrand-Berthelot (Lead / Corresponding author), Jose Palacios (Lead / Corresponding author)

    Research output: Contribution to journalArticlepeer-review

    12 Citations (Scopus)


    Bacteria require nickel transporters for the synthesis of Ni-containing metalloenzymes in natural, low nickel habitats. In this work we carry out functional and topological characterization of Rhizobium leguminosarum HupE, a nickel permease required for the provision of this element for [NiFe] hydrogenase synthesis. Expression studies in the Escherichia coli nikABCDE mutant strain HYD723 revealed that HupE is a medium-affinity permease (apparent K<inf>m</inf> 227 ± 21 nM; V<inf>max</inf> 49 ± 21 pmol Ni<sup>2+</sup> min<sup>-1</sup> mg<sup>-1</sup> bacterial dry weight) that functions as an energy-independent diffusion facilitator for the uptake of Ni(ii) ions. This Ni<sup>2+</sup> transport is not inhibited by similar cations such as Mn<sup>2+</sup>, Zn<sup>2+</sup>, or Co<sup>2+</sup>, but is blocked by Cu<sup>2+</sup>. Analysis of site-directed HupE mutants allowed the identification of several residues (H36, D42, H43, F69, E90, H130, and E133) that are essential for HupE-mediated Ni uptake in E. coli cells. By using translational fusions to reporter genes we demonstrated the presence of five transmembrane domains with a periplasmic N-terminal domain and a C-terminal domain buried in the lipid bilayer. The periplasmic N-terminal domain contributes to stability and functionality of the protein.

    Original languageEnglish
    Pages (from-to)691-701
    Number of pages11
    Issue number4
    Early online date28 Jan 2015
    Publication statusPublished - Apr 2015

    ASJC Scopus subject areas

    • Biomaterials
    • Metals and Alloys
    • Chemistry (miscellaneous)
    • Biochemistry
    • Biophysics


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