Abstract
Salmonella enterica serovar Typhimurium (S. Typhimurium) can survive in the phagosome of macrophages, causing serious medical and veterinary problems. CueP is uniquely found in S. Typhimurium and has been characterized as a major periplasmic copper-binding protein. Although cueP has been identified as being responsible for the copper resistance of the bacterium in vivo, the biochemical role and three-dimensional structure of CueP remain unknown. In this study, CueP from S. Typhimurium was overexpressed and the recombinant protein was purified using Ni-NTA affinity, anion-exchange and gel-filtration chromatographies. The purified CueP protein was crystallized using the vapour-diffusion method. A diffraction data set was collected to 2.5 Å resolution at 100 K. The crystal belonged to space group P2(1)2(1)2(1). To obtain initial phases, selenomethionyl-substituted protein was overproduced and purified. Optimization of crystallization conditions for the selenomethionyl-substituted protein is in progress.