Experimental Phasing Using Zinc and Sulfur Anomalous Signals Measured at the Zinc Absorption Peak SCIE SCOPUS KCI

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Title
Experimental Phasing Using Zinc and Sulfur Anomalous Signals Measured at the Zinc Absorption Peak
Author(s)
Lee, Sangmin; Kim, Min-Kyu; Ji, Chang-Jun; Lee, Jin-Won; Cha, Sun-Shin
Publication Year
2013-10
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 angstrom was collected at the zinc peak wavelength (lambda=1.2823 angstrom). 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.
ISSN
1225-8873
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/3092
DOI
10.1007/s12275-013-3412-2
Bibliographic Citation
JOURNAL OF MICROBIOLOGY, v.51, no.5, pp.639 - 643, 2013
Publisher
MICROBIOLOGICAL SOCIETY KOREA
Subject
AUTOMATED STRUCTURE SOLUTION; CRYSTAL-STRUCTURE; IRON HOMEOSTASIS; FUR; EXPRESSION; PROTEIN; GENE; TRANSCRIPTION; BINDING; SYSTEM
Keywords
Ferric uptake regulator; transcription regulator; crystallization; experimental phasing
Type
Article
Language
English
Document Type
Article
Publisher
MICROBIOLOGICAL SOCIETY KOREA
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