Abstract
Iron is an essential transition metal required for bacterial growth and survival. Excess free iron can lead to the generation of reactive oxygen species that can cause severe damage to cellular functions. Cells have developed iron-sensing regulators to maintain iron homeostasis at the transcription level. The ferric uptake regulator (Fur) is an iron-responsive regulator that controls the expression of genes involved in iron homeostasis, bacterial virulence, stress resistance, and redox metabolism. Here, we report the expression, purification, crystallization, and phasing of the apo-form of Bacillus subtilis Fur (BsFur) in the absence of regulatory metal ions. Crystals were obtained by microbatch crystallization method at 295 K and diffraction data at a resolution of 2.6 Å was collected at the zinc peak wavelength (λ=1.2823 Å). Experimental phasing identified the positions of one zinc atom and four sulfur atoms of cysteine residues coordinating the zinc atom, indicating that the data contained a meaningful anomalous scattering originating from the ordered zinc-coordinating sulfur atoms, in spite of the small anomalous signals of sulfur atoms at the examined wavelength.
Original language | English |
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Pages (from-to) | 639-643 |
Number of pages | 5 |
Journal | Journal of Microbiology |
Volume | 51 |
Issue number | 5 |
DOIs | |
State | Published - Oct 2013 |
Bibliographical note
Funding Information:This work was supported by the National Research Foundation of Korea Grant NRF-2012R1A2A2A02005978, the CAP through Korea Research Council of Fundamental Science Technology (KRCF), Korea Institute of Science and Technology (KIST), & Korea Institute of Ocean Science and Technology (KIOST), the Marine and Extreme Genome Research Center program, and the Development of Biohydrogen Production Technology Using Hyperthermophilic Archaea program of MOF. Works performed at Hanyang University were supported by the National Research Foundation of Korea Grant KRF-2008-313-C0077.
Keywords
- crystallization
- experimental phasing
- Ferric uptake regulator
- transcription regulator