Purification, crystallization, and preliminary X-ray crystallographic analysis of VV2_1132, a LysR-type transcriptional regulator from Vibrio vulnificus

• 발행기관: 한국구조생물학회
• 저자명: Yongdae Jang Garam Choi Inseong Jo Sang Ho Choi Nam-Chul Ha
• 간행물 정보: 『Biodesign』Vol 5, No 1, Mar, 44~48쪽, 전체 5쪽
• 주제분류: 자연과학 > 생물학
• 파일형태: PDF
• 발행일자: 2017.03.30

영문 초록

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.

목차

INTRODUCTION

RESULTS AND DISCUSSION

METHODS