Abstract

Article info 2017 5 (1)    005  (01)   pp.  44 ~ 48
Title Purification, crystallization, and preliminary X-ray crystallographic analysis of VV2_1132, a LysR-type transcriptional regulator from Vibrio vulnificus
Authors Yongdae Jang 1 , Garam Choi 1,2 , Inseong Jo 1 , Sang Ho Choi 1,2 and Nam-Chul Ha 1, *
Institutions 1 Department of Agricultural Biotechnology, Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea, 2 National Research Laboratory of Molecular Microbiology and Toxicology. *Correspondence: hanc210@snu.ac.kr
Abstract LysR-type transcriptional regulators (LTTRs) belong to the largest family of transcriptional regulators in prokaryotes. However, the crystal structures of only a few full-length LTTRs have been determined. LTTR HypT (also known as YjiE or QseD) from Escherichia coli specifically senses hypochlorite to protect oxidation damage by hypochlorite generated by the host immune system. In the genome of the highly virulent bacteria, Vibrio vulnificus, VV2_1132 showed the highest sequence similarity to E. coli HypT. In this study, we overexpressed full-length VV2_1132 in an E. coli expression system and crystallized the protein. The crystal diffracted X-rays to 2.2 Å resolution and belonged to the orthogonal space group P2 1 2 1 2, with unit cell parameters a = 57.8, b = 113.5, and c = 220.7 Å. Cell content analysis predicted that the asymmetric unit may contain four molecules of VV2_1132. For the case of four molecules in the asymmetric unit, the Matthews coefficient was 2.70 Å 3 /Da with a solvent content of 54.5%. We are currently identifying the crystal structure of VV2_1132 using anomalous signals from selenomethione-substituted crystals. This crystal structure will help elucidate the function and action mechanism of VV2_1132, which may be involved in the pathogenesis of V. vulnificus.