Novel protein engineering strategy for creating highly receptor-selective mutant TNFs |
| |
Authors: | Tetsuya Nomura Yasuhiro Abe Haruhiko Kamada Masaki Inoue Shuhei Arita Takeshi Furuya Yasuo Yoshioka Hiroko Shibata Takuya Yamashita Kazuya Nagano Yohei Mukai Madoka Taniai Shin-ichi Tsunoda Yasuo Tsutsumi |
| |
Institution: | a National Institute of Biomedical Innovation (NiBio), 7-6-8 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan b Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan c The Center for Advanced Medical Engineering and Informatics, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan d Hayashibara Biochemical Laboratories Inc., 675-1 Fujisaki, Okayama 702-8006, Japan |
| |
Abstract: | Tumor necrosis factor (TNF) plays important roles in host defense and in preventing tumor formation by acting via its receptors, TNFR1 and TNFR2, functions of which are less understood. To this end, we have been isolating TNF receptor-selective mutants using phage display technique. However, generation of a phage library with large repertoire (>108) is impeded by the limited transformation efficiency of Escherichia coli. Therefore, it is currently difficult to create a mutant library containing amino acid substitutions in more than seven residues. To overcome this problem, here we have used two different TNF mutant libraries, each containing random substitutions at six selected amino acid residues, and utilized a gene shuffling method to construct a randomized mutant library containing substitutions at 12 different amino acid residues of TNF. Consequently, using this library, we identified TNF mutants with greater receptor-selectivity and enhanced receptor-specific bioactivity than the existing mutants. |
| |
Keywords: | Tumor necrosis factor (TNF) Gene shuffling Phage display technique Protein engineering TNF receptor-selective mutant |
本文献已被 ScienceDirect 等数据库收录! |
|