A Pyrrolysine Analogue for Protein Click Chemistry
A Pyrrolysine Analogue for Protein Click Chemistry
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DOI：
10.1002/anie.200805420
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发表时间：
2009-01-01
影响因子：
16.6
通讯作者：
Chan, Michael K.
中科院分区：
文献类型：
--
作者：
Fekner, Tomasz;Li, Xin;Chan, Michael K.
忽略停止标志：通过UAG密码子，将含有末端炔的pyrlysine类似物位点特异性地结合到重组钙调蛋白（CaM）中。用含叠氮化物的染料用铜(I)催化的咔嗒反应对所得蛋白进行标记。随后应用正交半胱氨酸标记方法产生了一个CaM标记与两个不同的荧光团，使其研究FRET光谱。
Ignoring the STOP sign: A pyrrolysine analogue bearing a terminal alkyne was site‐specifically incorporated into recombinant calmodulin (CaM) through a UAG codon. The resulting protein was labeled with an azide‐containing dye using a copper(I)‐catalyzed click reaction. Subsequent application of an orthogonal cysteine tagging method yielded a CaM labeled with two distinct fluorophores that enabled its study by FRET spectroscopy.