Spontaneous Orthogonal Protein Crosslinking via a Genetically Encoded 2-Carboxy-4-Aryl-1,2,3-Triazole.
Spontaneous Orthogonal Protein Crosslinking via a Genetically Encoded 2-Carboxy-4-Aryl-1,2,3-Triazole.
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DOI：
10.1002/anie.202202657
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发表时间：
2022-05-23
影响因子：
16.6
通讯作者：
Lin, Qing
中科院分区：
文献类型：
--
作者：
Xu, Yali;Rahim, Abdur;Lin, Qing
关键词：
在这里，我们报道了N_2-羧基-4-芳基-1，2，3-三氮唑赖氨酸(CATKs)的设计及其通过遗传密码扩展的位点特异性结合到蛋白质中。当被引入蛋白质二聚体界面时，CATKs允许自发的、邻近驱动的、位置选择性的交联在活细胞中产生共价蛋白质二聚体，含苯基的CATK-1对近端的Lys和Tyr显示出高的反应性。此外，当将CATK-1引入单链VHH抗体或增压单体的N端β链中时，CATK-1能够实现位点特异性的链间、与位于相反β链上的近端Tyr的正交交联。与未交联单体膜相比，正交交联单体膜显示出更好的细胞摄取和对内体酶的蛋白分解稳定性。CATKs强大的交联性有助于设计具有改进的物理化学性质的新的蛋白质拓扑结构。基因编码的N_2-羧基-4-芳基-1，2，3-三氮唑-赖氨酸(CATK)允许自发的、定量的、位点特异性的链间交联与小抗体模拟蛋白中的近端Tyr，产生一种新的蛋白质拓扑结构，伴随着改善的细胞摄取和增强的蛋白水解性。
Here we report the design of N2-carboxy-4-aryl-1,2,3-triazole-lysines (CATKs) and their site-specific incorporation into proteins via genetic code expansion. When introduced into the protein dimer interface, CATKs permitted spontaneous, proximity-driven, site-selective crosslinking to generate covalent protein dimers in living cells, with phenyl-bearing CATK-1 exhibiting high reactivity toward the proximal Lys and Tyr. Furthermore, when introduced into the N-terminal β-strand of either a single-chain VHH antibody or a supercharged monobody, CATK-1 enabled site-specific, inter-strand, orthogonal crosslinking with a proximal Tyr located on the opposing β-strand. Compared with a non-crosslinked monobody, the orthogonally crosslinked monobody displayed improved cellular uptake and enhanced proteolytic stability against an endosomal enzyme. The robust crosslinking reactivity of CATKs should facilitate the design of novel protein topologies with improved physicochemical properties. A genetically encoded N2-carboxy-4-aryl-1,2,3-triazole-lysine (CATK) permits a spontaneous, quantitative, site-specific inter-strand crosslinking with a proximal Tyr in a small antibody-mimetic protein, generating a novel protein topology accompanied by improved cellular uptake and enhanced proteolytic resistance.
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DOI：
10.1002/anie.201308794
发表时间：
2014-02-17
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Xiang Z;Lacey VK;Ren H;Xu J;Burban DJ;Jennings PA;Wang L
通讯作者：
Wang L
影响因子：
5.6
作者：
Saerens, Dirk;Conrath, Katja;Muyldermans, Serge
通讯作者：
Muyldermans, Serge
影响因子：
4.8
作者：
Govaert, Jochen;Pellis, Mireille;Saerens, Dirk
通讯作者：
Saerens, Dirk
DOI：
10.1002/pro.3148
发表时间：
2017-05
期刊：
Protein science : a publication of the Protein Society
影响因子：
--
作者：
Sha F;Salzman G;Gupta A;Koide S
通讯作者：
Koide S
影响因子：
14.8
作者：
Adibekian, Alexander;Martin, Brent R.;Wang, Chu;Hsu, Ku-Lung;Bachovchin, Daniel A.;Niessen, Sherry;Hoover, Heather;Cravatt, Benjamin F.
通讯作者：
Cravatt, Benjamin F.