Affinity maturation of Cry1Aa toxin to the Bombyx mori cadherin-like receptor by directed evolution based on phage display and biopanning selections of domain II loop 2 mutant toxins.
Affinity maturation of Cry1Aa toxin to the Bombyx mori cadherin-like receptor by directed evolution based on phage display and biopanning selections of domain II loop 2 mutant toxins.
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DOI：
10.1002/mbo3.188
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发表时间：
2014-08
期刊：
影响因子：
3.4
通讯作者：
Sato, Ryoichi
中科院分区：
文献类型：
--
作者：
Endo, Haruka;Kobayashi, Yuki;Hoshino, Yasushi;Tanaka, Shiho;Kikuta, Shingo;Tabunoki, Hiroko;Sato, Ryoichi
利用噬菌体展示和生物扫描技术定向进化Cry1Aa毒素，以产生更高的与家蚕钙粘素样受体(BtR175)的结合亲和力。从含有BtR175结构域II环2突变毒素的噬菌体文库中筛选出与BtR175结合亲和力分别高16倍、16倍和50倍的3种突变体毒素(371WGLA374、371WPHH374和371WRPQ37425)。然而，这3种突变体对家蚕幼虫和表达BtR175毒素结合区的培养细胞的观察到的毒力并没有增加，这表明与钙粘附素结合亲和力的增加并不有助于杀虫活性。Cry毒素通过定向进化对受体的亲和力成熟相对容易实现，并且似乎有可能产生具有更高杀虫活性的毒素。
Directed evolution of a Cry1Aa toxin using phage display and biopanning was performed to generate an increased binding affinity to the Bombyx mori cadherin-like receptor (BtR175). Three mutant toxins (371WGLA374, 371WPHH374, 371WRPQ37425) with 16-, 16-, and 50-fold higher binding affinities, respectively, for BtR175 were selected from a phage library containing toxins with mutations in domain II loop 2. However, the observed toxicities of the three mutants against B. mori larvae and cultured cells expressing the BtR175 toxin-binding region did not increase, suggesting that increased binding affinity to cadherins does not contribute to the insecticidal activity. Affinity maturation of a Cry toxin to a receptor via directed evolution was relatively simple to achieve, and seems to have potential for generating a toxin with increased insecticidal activity.
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影响因子：
4.5
作者：
Gahan LJ;Pauchet Y;Vogel H;Heckel DG
通讯作者：
Heckel DG
影响因子：
3
作者：
Elena Fernandez, Luisa;Gomez, Isabel;Soberon, Mario
通讯作者：
Soberon, Mario
影响因子：
2.4
作者：
Liu, XS;Dean, DH
通讯作者：
Dean, DH
DOI：
10.1073/pnas.0831036100
发表时间：
2003-04-29
影响因子：
11.1
作者：
Morin, S;Biggs, RW;Tabashnik, BE
通讯作者：
Tabashnik, BE
影响因子：
4
作者：
Fujii Y;Tanaka S;Otsuki M;Hoshino Y;Morimoto C;Kotani T;Harashima Y;Endo H;Yoshizawa Y;Sato R
通讯作者：
Sato R