Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic Lacinutrix jangbogonensis PAMC 27137

Crystals

T. Doohun Kim¹  Aekyung Park²  Hongshi Jin³  Seung Chul Shin³  Jonghyeon Son⁴  Jun Hyuck Lee⁵  Woong Choi⁵  Han-Woo Kim⁵ 

[1] Department of Chemistry, Sookmyung Women’s University, Seoul 04310, Korea;Division of Emerging Infectious Diseases Bureau of Infectious Disease Diagnosis Control, Korea Disease Control and Prevention Agency, Cheongju 28159, Korea;Division of Life Sciences, Korea Polar Research Institute, Incheon 21990, Korea;New Drug Development Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu 41061, Korea;Research Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, Korea;

关键词: single stranded DNA binding protein;    Lacinutrix jangbogonensis PAMC 27137;    X-ray crystallography;   

DOI  :  10.3390/cryst12040538

来源: DOAJ

【 摘 要 】

Single-stranded DNA-binding proteins (SSBs) are essential for DNA metabolism, including repair and replication, in all organisms. SSBs have potential applications in molecular biology and in analytical methods. In this study, for the first time, we purified, structurally characterized, and analyzed psychrophilic SSB (LjSSB) from Lacinutrix jangbogonensis PAMC 27137 isolated from the Antarctic region. LjSSB has a relatively short amino acid sequence, consisting of 111 residues, with a molecular mass of 12.6 kDa. LjSSB protein was overexpressed in Escherichia coli BL21 (DE3) and analyzed for binding affinity using 20- and 35-mer deoxythymidine oligonucleotides (dT). In addition, the crystal structure of LjSSB at a resolution 2.6 Å was obtained. The LjSSB protein crystal belongs to the space group C222 with the unit cell parameters of a = 106.58 Å, b = 234.14 Å, c = 66.14 Å. The crystal structure was solved using molecular replacement, and subsequent iterative structure refinements and model building are currently under progress. Further, the complete structural information of LjSSB will provide a novel strategy for protein engineering and for the application on molecular biological techniques.

【 授权许可】

Unknown