Creation of mouse TNFR2-selective agonistic TNF mutants using a phage display technique.
Creation of mouse TNFR2-selective agonistic TNF mutants using a phage display technique.
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DOI：
10.1016/j.bbrep.2016.06.008
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发表时间：
2016-09
影响因子：
2.7
通讯作者：
Tsunoda SI
中科院分区：
文献类型：
--
作者：
Ando D;Inoue M;Kamada H;Taki S;Furuya T;Abe Y;Nagano K;Tsutsumi Y;Tsunoda SI
肿瘤坏死因子-α (TNF)是一种免疫调节细胞因子，已被认为通过结合两种类型的TNF受体（TNFR1/TNFR2）引起炎症反应并保护组织功能障碍。然而，tnfr2特异性激活的生理效应尚不清楚。因此，我们旨在产生具有完全tnfr2选择性激动剂活性的TNF突变体作为功能分析工具。在本研究中，我们利用噬菌体展示技术创建小鼠TNFR2 （mTNFR2）选择性TNF突变体，该突变体特异性结合mTNFR2，并与野生型TNF相比显示出充分的生物活性。建立了一个新的噬菌体文库，显示TNF突变体，其中预测受体结合位点的9个氨基酸残基随机化。从这个文库中，分离出一个对mTNFR2具有高结合选择性和生物活性的激动性TNF突变体。我们认为这种TNF突变体将是阐明mTNFR2功能作用的有力工具。我们产生了一个具有完全tnfr2选择性激动剂活性的TNF突变体。该突变体是用噬菌体展示技术鉴定的。该激动剂对小鼠TNFR2具有高结合选择性和生物活性。这将是阐明小鼠TNFR2功能作用的有力工具。
Tumor necrosis factor-α (TNF), which is an immuno-modulatory cytokine, has been suggested to cause inflammatory responses as well as protection against tissue dysfunction by binding two types of TNF receptor (TNFR1/TNFR2). However, the physiological effects of TNFR2-specific activation remain unclear. We therefore aimed to generate a TNF mutant with full TNFR2-selective agonist activity as a functional analytical tool. In this study, we utilized a phage display technique to create mouse TNFR2 (mTNFR2)-selective TNF mutants that bind specifically to mTNFR2 and show full bioactivity compared with wild-type TNF. A new phage library displaying TNF mutants was created, in which nine amino acid residues at the predicted receptor-binding site were randomized. From this library, an agonistic TNF mutant exhibiting high binding selectivity and bioactivity to mTNFR2 was isolated. We propose that this TNF mutant would be a powerful tool with which to elucidate the functional roles of mTNFR2. We generated a TNF mutant with full TNFR2-selective agonist activity. This mutant was identified using a phage display technique. This agonist exhibited high binding selectivity and bioactivity to mouse TNFR2. This would be a powerful tool to elucidate the functional roles of mouse TNFR2.