Residue choice defines efficiency and influence of bioorthogonal protein modification via genetically encoded strain promoted Click chemistry
Residue choice defines efficiency and influence of bioorthogonal protein modification via genetically encoded strain promoted Click chemistry
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DOI：
10.1039/c2cc31887c
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发表时间：
2012-01-01
影响因子：
4.9
通讯作者：
Jones, D. Dafydd
中科院分区：
文献类型：
--
作者：
Reddington, Samuel C.;Tippmann, Eric M.;Jones, D. Dafydd
GFP和FRET兼容染料用于评估遗传编码的叠氮化物定位对单击化学基于化学的蛋白结合的影响。尽管使用菌株促进反应的采样突变体的修改效率与10倍相似，但与GFP表面上的芳基叠氮化物的可及性没有简单的相关性。一个标记的GFP突变体（GLN204AZPHE）表现出高效率的FRET（类似于90％），并且前所未有的伪stokes偏移为126 nm。
GFP and a FRET compatible dye were used to assess the influence of genetically encoded aryl azide positioning on Click chemistry-based protein conjugation. While modification efficiency of the sampled mutants using a strain promoted reaction varied by as much as similar to 10 fold, there was no simple correlation with accessibility of the aryl azide on GFP's surface. One labeled GFP mutant (Gln204AzPhe) exhibited high efficiency FRET (similar to 90%) and an unprecedented pseudo-Stokes shift of 126 nm.