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Experimental phasing using zinc and sulfur anomalous signals measured at the zinc absorption peak.
- Source :
-
Journal of microbiology (Seoul, Korea) [J Microbiol] 2013 Oct; Vol. 51 (5), pp. 639-43. Date of Electronic Publication: 2013 Oct 31. - Publication Year :
- 2013
-
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.
- Subjects :
- Bacillus subtilis isolation & purification
Bacterial Proteins isolation & purification
Crystallography, X-Ray
Gene Expression
Models, Molecular
Repressor Proteins isolation & purification
Spectrum Analysis
Bacillus subtilis chemistry
Bacterial Proteins chemistry
Repressor Proteins chemistry
Sulfur analysis
Zinc analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1976-3794
- Volume :
- 51
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of microbiology (Seoul, Korea)
- Publication Type :
- Academic Journal
- Accession number :
- 24173644
- Full Text :
- https://doi.org/10.1007/s12275-013-3412-2