Crystallization and preliminary X-ray crystallographic analysis of the Escherichia coli outer membrane cobalamin transporter BtuB in complex with the carboxy-terminal domain of TonB
Abstract
Crystals of a complex of the E. coli proteins BtuB (outer membrane cobalamin transporter) and TonB (carboxy-terminal domain) diffracting to 2.1 Å resolution have been obtained. The energy-dependent uptake of organometallic compounds and other micronutrients across the outer membranes of Gram-negative bacteria is carried out by outer membrane active-transport proteins that utilize the proton-motive force of the inner membrane via coupling to the TonB protein. The Escherichia coli outer membrane cobalamin transporter BtuB and a carboxy-terminal domain of the TonB protein, residues 147–239 of the wild-type protein, were expressed and purified individually. A complex of BtuB and TonB{sup 147–239} was formed in the presence of the substrate cyanocobalamin (CN-Cbl; vitamin B{sub 12}) and calcium and was crystallized. BtuB was purified in the detergent LDAO (n-dodecyl-N,N-dimethylamine-N-oxide) and the complex was formed in a detergent mixture of LDAO and C{sub 8}E{sub 4} (tetraethylene glycol monooctylether). Crystals were obtained by sitting-drop vapor diffusion, with the reservoir containing 30%(v/v) polyethylene glycol (PEG 300) and 100 mM sodium acetate pH 5.2. The crystals belong to space group P2{sub 1}2{sub 1}2{sub 1} (unit-cell parameters a = 74.3, b = 82.4, c = 122.6 Å). The asymmetric unit consists of a single BtuB–TonB complex. Data sets havemore »
- Authors:
-
- Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22908 (United States)
- Interdisciplinary Graduate Program in Biophysics, University of Virginia, Charlottesville, Virginia 22908 (United States)
- Publication Date:
- OSTI Identifier:
- 22360210
- Resource Type:
- Journal Article
- Journal Name:
- Acta Crystallographica. Section F
- Additional Journal Information:
- Journal Volume: 62; Journal Issue: Pt 7; Other Information: PMCID: PMC2242962; PMID: 16820681; PUBLISHER-ID: pu5141; OAI: oai:pubmedcentral.nih.gov:2242962; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; ACETATES; CALCIUM; COUPLING; CRYSTALLIZATION; CRYSTALS; DIFFUSION; ESCHERICHIA COLI; MEMBRANES; MIXTURES; OXIDES; RESOLUTION; SODIUM; SPACE GROUPS; SUBSTRATES
Citation Formats
Shultis, David D., Purdy, Michael D., Banchs, Christian N., Wiener, Michael C., E-mail: mwiener@virginia.edu, and Interdisciplinary Graduate Program in Biophysics, University of Virginia, Charlottesville, Virginia 22908. Crystallization and preliminary X-ray crystallographic analysis of the Escherichia coli outer membrane cobalamin transporter BtuB in complex with the carboxy-terminal domain of TonB. United Kingdom: N. p., 2006.
Web. doi:10.1107/S1744309106018240.
Shultis, David D., Purdy, Michael D., Banchs, Christian N., Wiener, Michael C., E-mail: mwiener@virginia.edu, & Interdisciplinary Graduate Program in Biophysics, University of Virginia, Charlottesville, Virginia 22908. Crystallization and preliminary X-ray crystallographic analysis of the Escherichia coli outer membrane cobalamin transporter BtuB in complex with the carboxy-terminal domain of TonB. United Kingdom. https://doi.org/10.1107/S1744309106018240
Shultis, David D., Purdy, Michael D., Banchs, Christian N., Wiener, Michael C., E-mail: mwiener@virginia.edu, and Interdisciplinary Graduate Program in Biophysics, University of Virginia, Charlottesville, Virginia 22908. 2006.
"Crystallization and preliminary X-ray crystallographic analysis of the Escherichia coli outer membrane cobalamin transporter BtuB in complex with the carboxy-terminal domain of TonB". United Kingdom. https://doi.org/10.1107/S1744309106018240.
@article{osti_22360210,
title = {Crystallization and preliminary X-ray crystallographic analysis of the Escherichia coli outer membrane cobalamin transporter BtuB in complex with the carboxy-terminal domain of TonB},
author = {Shultis, David D. and Purdy, Michael D. and Banchs, Christian N. and Wiener, Michael C., E-mail: mwiener@virginia.edu and Interdisciplinary Graduate Program in Biophysics, University of Virginia, Charlottesville, Virginia 22908},
abstractNote = {Crystals of a complex of the E. coli proteins BtuB (outer membrane cobalamin transporter) and TonB (carboxy-terminal domain) diffracting to 2.1 Å resolution have been obtained. The energy-dependent uptake of organometallic compounds and other micronutrients across the outer membranes of Gram-negative bacteria is carried out by outer membrane active-transport proteins that utilize the proton-motive force of the inner membrane via coupling to the TonB protein. The Escherichia coli outer membrane cobalamin transporter BtuB and a carboxy-terminal domain of the TonB protein, residues 147–239 of the wild-type protein, were expressed and purified individually. A complex of BtuB and TonB{sup 147–239} was formed in the presence of the substrate cyanocobalamin (CN-Cbl; vitamin B{sub 12}) and calcium and was crystallized. BtuB was purified in the detergent LDAO (n-dodecyl-N,N-dimethylamine-N-oxide) and the complex was formed in a detergent mixture of LDAO and C{sub 8}E{sub 4} (tetraethylene glycol monooctylether). Crystals were obtained by sitting-drop vapor diffusion, with the reservoir containing 30%(v/v) polyethylene glycol (PEG 300) and 100 mM sodium acetate pH 5.2. The crystals belong to space group P2{sub 1}2{sub 1}2{sub 1} (unit-cell parameters a = 74.3, b = 82.4, c = 122.6 Å). The asymmetric unit consists of a single BtuB–TonB complex. Data sets have been collected to 2.1 Å resolution at a synchrotron beamline (APS SER-CAT 22-ID)},
doi = {10.1107/S1744309106018240},
url = {https://www.osti.gov/biblio/22360210},
journal = {Acta Crystallographica. Section F},
issn = {1744-3091},
number = Pt 7,
volume = 62,
place = {United Kingdom},
year = {Sat Jul 01 00:00:00 EDT 2006},
month = {Sat Jul 01 00:00:00 EDT 2006}
}