Experimental phasing using zinc and sulfur anomalous signals measured at the zinc absorption peak

Sangmin Lee, Min Kyu Kim, Chang Jun Ji, Jin Won Lee, Sun Shin Cha

Research output: Contribution to journalArticlepeer-review


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 languageEnglish
Pages (from-to)639-643
Number of pages5
JournalJournal of Microbiology
Issue number5
StatePublished - Oct 2013


  • crystallization
  • experimental phasing
  • Ferric uptake regulator
  • transcription regulator


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