|
KMID : 0880220130510050639
|
|
Journal of Microbiology
2013 Volume.51 No. 5 p.639 ~ p.643
|
|
|
Experimental Phasing Using Zinc and Sulfur Anomalous Signals Measured at the Zinc Absorption Peak
|
|
Lee Sang-Min
Kim Min-Kyu
Ji Chang-Jun
Lee Jin-Won
Cha Sun-Shin
|
|
|
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 A was collected at the zinc peak wavelength (λ=1.2823 A). 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.
|
|
|
KEYWORD
|
|
|
Ferric uptake regulator, transcription regulator, crystallization, experimental phasing
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|