Crystallization and preliminary X-ray diffraction analysis of a soluble domain of the putative zinc transporter CzrB from Thermus thermophilus

Journal Article · Wed Aug 01 04:00:00 EDT 2007 · Acta Crystallographica. Section F

DOI:https://doi.org/10.1107/S1744309107032277· OSTI ID:22360389

Höfer, Nicole; Kolaj, Olga; Li, Hui ^[1]; Cherezov, Vadim ^[2]; Gillilan, Richard ^[3]; Wall, J. Gerard ^[1]; Caffrey, Martin ^[1]

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1. Department of Chemical and Environmental Sciences and Materials and Surface Science Institute, University of Limerick, Limerick (Ireland)
2. Department of Chemistry, The Ohio State University, Columbus, OH 43210 (United States)
3. Cornell High Energy Synchrotron Source, Cornell University, Ithaca, NY 14850 (United States)

Diffraction-quality crystals of the apo (1.7 Å) and zinc-bound forms (2.2 Å) of the water-soluble C-terminal domain of the putative zinc transporter CzrB from T. thermophilus have been grown using recombinant production of the protein in E. coli and a combination of vapour-diffusion, batch and seeding crystallogenesis techniques. CzrB is a putative zinc transporter from Thermus thermophilus. The protein is proposed to consist of a hexahelical transmembrane domain with a cytosolic extramembranal C-terminus. The latter 92-residue fragment may be expressed free and may function independently of the full-length integral membrane protein. A 6×His-tagged form of the water-soluble fragment has been overexpressed in Escherichia coli and diffraction-quality crystals of the tagged and tag-free variants have been grown. Preliminary X-ray analyses of tag-free fragment crystals with (2.2 Å resolution) and without zinc ions (1.7 Å resolution) reveal that the former has at least two zinc ions bound per monomer.

• OSTI ID: 22360389
• Journal Information: Acta Crystallographica. Section F, Journal Name: Acta Crystallographica. Section F Journal Issue: Pt 8 Vol. 63; ISSN ACSFCL; ISSN 1744-3091
• Country of Publication: United Kingdom
• Language: English

Related Subjects

75 CONDENSED MATTER PHYSICS
SUPERCONDUCTIVITY AND SUPERFLUIDITY
CRYSTALLIZATION
CRYSTALS
DIFFUSION
ESCHERICHIA COLI
LENGTH
MEMBRANE PROTEINS
MONOMERS
RESOLUTION
WATER
X-RAY DIFFRACTION
ZINC
ZINC IONS