|JKMRS Volume 17, No 1 30-39; Expression and Purification of Un...|
|2013년 06월 20일 / 조회수: 558|
Expression and Purification of Unstructured Protein, IMUP-1, using Chaperone Co-expression System for NMR Study
Jong-Jae Yi, Jung Ki Yoo, Jin Kyeoung Kim, and Woo Sung Son*
College of pharmacy, CHA University, 120 Haeyong-ro, Pocheon-si, Gyeonggi-do, 487-010, Republic of Korea
Received March 21, 2013; Revised June 4, 2013; Accepted June 10, 2013
Abstract Immortalization-upregulated protein-1 (IMUP-1) genes have been cloned and are known to be involved in SV40-mediated immortalization. IMUP-1 gene is highly expressed in various cancer cell lines and tumors, suggesting the possibility that they might be involved in tumorigenicity. Previously, there were several problems for overexpression of IMUP-1 in bacterial expression systems including low solubility and aggregation due to unstructured property. To investigate the structural properties, it is necessary to obtain lots of pure and soluble proteins. Accordingly, the co-expression systems of bacterial chaperone proteins, GroEL-GroES, were used to increase solubility of IMUP-1. From the analysis of NMR and CD experiment data, it is suggested that the protein adopt typical the random coil properties in solution.
Keywords IMUP-1, Chaperone, Purification, CD, NMR