Structure and Metal Binding Properties of ZnuA, a Periplasmic Zinc Transporter from Escherichia coli
Document Type
Article
Language
eng
Format of Original
18 p.
Publication Date
2-2008
Publisher
Springer
Source Publication
Journal of Biological Inorganic Chemistry
Source ISSN
0949-8257
Original Item ID
DOI: 10.1007/s00775-007-0320-0, PubMed Central: PMC2630496
Abstract
ZnuA is the periplasmic Zn2+-binding protein associated with the high-affinity ATP-binding cassette ZnuABC transporter from Escherichia coli. Although several structures of ZnuA and its homologs have been determined, details regarding metal ion stoichiometry, affinity, and specificity as well as the mechanism of metal uptake and transfer remain unclear. The crystal structures of E. coli ZnuA (Eco-ZnuA) in the apo, Zn2+-bound, and Co2+-bound forms have been determined. ZnZnuA binds at least two metal ions. The first, observed previously in other structures, is coordinated tetrahedrally by Glu59, His60, His143, and His207. Replacement of Zn2+ with Co2+ results in almost identical coordination geometry at this site. The second metal binding site involves His224 and several yet to be identified residues from the His-rich loop that is unique to Zn2+ periplasmic metal binding receptors. Electron paramagnetic resonance and X-ray absorption spectroscopic data on CoZnuA provide additional insight into possible residues involved in this second site. The second site is also detected by metal analysis and circular dichroism (CD) titrations. Eco-ZnuA binds Zn2+ (estimated K d < 20 nM), Co2+, Ni2+, Cu2+, Cu+, and Cd2+, but not Mn2+. Finally, conformational changes upon metal binding observed in the crystal structures together with fluorescence and CD data indicate that only Zn2+ substantially stabilizes ZnuA and might facilitate recognition of ZnuB and subsequent metal transfer.
Recommended Citation
Yatsunyk, Liliya A.; Easton, J. Allen; Kim, Lydia R.; Sugarbaker, Stacy A.; Bennett, Brian; Breece, Robert M.; Vorontsov, Ivan I.; Tierney, David L.; Crowder, Michael W.; and Rosenzweig, Amy C., "Structure and Metal Binding Properties of ZnuA, a Periplasmic Zinc Transporter from Escherichia coli" (2008). Physics Faculty Research and Publications. 88.
https://epublications.marquette.edu/physics_fac/88
Comments
Accepted version. Journal of Biological Inorganic Chemistry, Vol. 13, No. 2 (February 2008): 271-288. DOI. © 2008 Springer. Used with permission.
The final publication is available at Springer via http://dx.doi.org/10.1007/s00775-007-0320-0.
Brian Bennett was affiliated with the Medical College of Wisconsin at the time of publication.
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