Synthesis and Incorporation of Unnatural Amino Acids To Probe and Optimize Protein Bioconjugations.
Synthesis and Incorporation of Unnatural Amino Acids To Probe and Optimize Protein Bioconjugations.
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DOI：
10.1021/acs.bioconjchem.5b00424
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发表时间：
2015-09-16
影响因子：
4.7
通讯作者：
Young DD
中科院分区：
文献类型：
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作者：
Maza JC;McKenna JR;Raliski BK;Freedman MT;Young DD
非天然氨基酸（UAA）在生物共轭中的利用是理想的，因为它们能够赋予一定程度的生物正交性和特异性。为了阐明制备 UAA 生物缀合物的最佳条件，我们合成了 9 种具有可变亚甲基键 (2-4) 和叠氮化物、炔烃或卤化物官能团的 UAA。然后使用混杂的氨酰基-tRNA 合成酶将所有 9 个 UAA 掺入绿色荧光蛋白 (GFP) 中。然后通过与荧光团或衍生树脂反应来分析不同的生物缀合以获得最佳系链长度。有趣的是，发现最佳系链长度取决于反应类型。总体而言，这些发现使人们更好地了解了可以优化以有效制备生物缀合物的各种参数。
The utilization of unnatural amino acids (UAAs) in bioconjugations is ideal due to their ability to confer a degree of bioorthogonality and specificity. In order to elucidate optimal conditions for the preparation of bioconjugates with UAAs, we synthesized 9 UAAs with variable methylene tethers (2–4) and either an azide, alkyne, or halide functional group. All 9 UAAs were then incorporated into green fluorescent protein (GFP) using a promiscuous aminoacyl-tRNA synthetase. The different bioconjugations were then analyzed for optimal tether length via reaction with either a fluorophore or a derivatized resin. Interestingly, the optimal tether length was found to be dependent on the type of reaction. Overall, these findings provide a better understanding of various parameters that can be optimized for the efficient preparation of bioconjugates.