crystallization communications Acta Cryst. (2011). F67, 1623–1626 doi:10.1107/S1744309111028995 1623 Acta Crystallographica Section F Structural Biology and Crystallization Communications ISSN 1744-3091 Cloning, purification and crystallographic analysis of a hypothetical protein, BPSL1549, from Burkholderia pseudomallei Abimael Cruz-Migoni, a Sergey N. Ruzheinikov, a Svetlana E. Sedelnikova, a Barbara Obeng, a Sylvia Chieng, b Rahmah Mohamed, b,c Sheila Nathan, b,c Patrick J. Baker a and David W. Rice a * a Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, The University of Sheffield, Sheffield S10 2TN, England, b School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor DE, Malaysia, and c Malaysia Genome Institute, UKM-MTDC Technology Centre, 43600 Bangi, Selangor DE, Malaysia Correspondence e-mail: d.rice@sheffield.ac.uk Received 20 June 2011 Accepted 18 July 2011 Burkholderia pseudomallei BPSL1549, a putative protein of unknown function, has been overexpressed in Escherichia coli, purified and subsequently crystal- lized by the hanging-drop vapour-diffusion method using PEG as a precipitant to give crystals with overall dimensions of 0.15  0.15  0.1 mm. Native data were collected to 1.47 A ˚ resolution at the European Synchrotron Radiation Facility (ESRF). The crystals belonged to space group P2 1 2 1 2 1 , with unit-cell parameters a = 37.1, b = 45.4, c = 111.9 A ˚ and with a single polypeptide chain in the asymmetric unit. 1. Introduction Burkholderia pseudomallei is a Gram-negative, facultative, intra- cellular pathogen of both animals and humans that grows in hot wet soil and stagnant water (Dance, 1991; Galyov et al. , 2010). This bacterium is the causative agent of melioidosis (Brett & Woods, 2000), a disease endemic to tropical and subtropical areas, particu- larly Southeast Asia and Northern Australia (Gan, 2005). Infection by B. pseudomallei occurs by inhalation, inoculation or ingestion of contaminated soil or water, with the size of the inoculum influencing the pattern and the severity of the disease (Cheng & Currie, 2005). Commonly, melioidosis has a complex spectrum of clinical manifes- tations, including abscesses, pneumonia and, at worst, fatal septi- caemia. The formation of abscesses can occur in any organ, although in patients with chronic melioidosis the lungs are commonly affected (Dhiensiri et al., 1988; Currie et al. , 2000). As a result of these symptoms mimicking those of other common diseases, such as malaria, typhoid fever, leptospirosis and tuberculosis (Leelarasamee & Bovornkitti, 1989), melioidosis is often misdiagnosed, making it difficult to treat in a timely fashion (Aldhous, 2005). There is currently no protective vaccine against melioidosis (Dance et al. , 1989; Bossi et al., 2004) and treatment requires intensive antibiotic therapy for many months. Moreover, many patients have a high risk of relapse after a dormant state of many decades (Chaowagul et al., 1993; Ngauy et al. , 2005) and the emergence of drug-resistant strains is reducing the efficacy of conventional therapies such as ceftazidime or imipenem (Simpson et al., 1999). The importance of B. pseudomallei as a disease-causing organism has now resulted in the completion of the sequencing of the genomes of a range of strains, including the most well characterized, K96243 (Holden et al., 2004). Genomic analysis has shown that B. pseudo- mallei contains two chromosomes of size 5.23 and 9.73 Mb with a high G + C content. The large chromosome contains a higher proportion of coding sequences involved in core functions associated with central metabolism and cell growth, whereas the small chromosome carries genes encoding accessory functions associated with adaptation and survival in different environments (Holden et al., 2004). Whilst some virulence factors involved in the pathogenesis of B. pseudomallei have been proposed (Adler et al., 2009; Galyov et al. , 2010), the molecular basis of the pathogenicity of the bacterium is poorly understood (Stone, 2007). Proteomic studies have led to the identi- fication of a number of proteins whose levels differ between the pathogenic Burkholderia strain B. pseudomallei and the nonpatho- genic strain B. thailandensis. These proteins therefore constitute a list # 2011 International Union of Crystallography All rights reserved