Expression, Purification and Characterization of Recombinant Toxins Consisting of Truncated Gastrin 17 and Pseudomonas Exotoxin

Title:Expression, Purification and Characterization of Recombinant Toxins Consisting of Truncated Gastrin 17 and Pseudomonas Exotoxin

Volume: 22 Issue: 2

Author(s): Xiao-Li Feng, Xi-Lin Liu, Shi-Ying Lu, Hong-Lin Ren, Yan-Song Li, Pan Hu, Quan Wang, Weihua Tong, Dong-Ming Yan, Yu Zhou, Song Zhang, Wen Jin and Zeng-Shan Liu

Affiliation:

Keywords: Cytotoxicity, expression, optimization, purification, truncated gastrin 17.

Abstract: Gastric cancer is a major cause of mortality and morbidity around world. However the effectiveness of the current approaches to the diagnosis and treatment of gastric cancer is limited. Recombinant targeted toxins may represent a novel direction of cancer therapy. In this study, we aimed to explore whether recombinant toxins fused with the truncated forms of G17 could target to kill cancer cells by recognizing CCK2R. Four recombinant Pseudomonas toxins PE38 fused with the forward or reverse truncated forms of G17 (G14 and G13) were successfully constructed, expressed, and purified. Their characteristics were further analyzed by SDS-PAGE, western blot and indirect immunofluorescence assay. The cytotoxicity assay demonstrated that only reversely fused recombinant toxins rG14PE38 and rG13PE38 exhibited certain toxicity on several cancer cell lines, and a competition assay indicated that the binding of the reverse gastrin-endotoxin to CCK2R (+) cells may be mediated by interaction between gastrin/gastrin-like and CCK2R.

Cite this article as:

Feng Xiao-Li, Liu Xi-Lin, Lu Shi-Ying, Ren Hong-Lin, Li Yan-Song, Hu Pan, Wang Quan, Tong Weihua, Yan Dong-Ming, Zhou Yu, Zhang Song, Jin Wen and Liu Zeng-Shan, Expression, Purification and Characterization of Recombinant Toxins Consisting of Truncated Gastrin 17 and Pseudomonas Exotoxin, Protein & Peptide Letters 2015; 22 (2) . https://dx.doi.org/10.2174/0929866521666141028214723