N6-(2-(R)-propargylglycyl)lysine as a clickable pyrrolysine mimic.
N6-(2-(R)-propargylglycyl)lysine as a clickable pyrrolysine mimic.
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DOI：
10.1002/asia.201000205
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发表时间：
2010-08-02
影响因子：
4.1
通讯作者：
Chan, Michael K.
中科院分区：
文献类型：
--
作者：
Li, Xin;Fekner, Tomasz;Chan, Michael K.
用有机荧光团对蛋白质进行定点标记[2]已被证明是研究广泛的生物系统结构和功能的一种有效方法。[3-5]这种方法最具挑战性的方面是在蛋白质链中合理选择的氨基酸残基上或其附近加入合适的荧光团。吡咯赖氨酸(1，图1)，第22个基因编码的氨基酸，可以结合到重组蛋白中以响应UAG密码子，[6，7]，这促使我们寻找具有适合锚定小的有机荧光团的反应功能的1的类似可结合类似物。我们以前报道过含有四氢呋喃的赖氨酸衍生物2的合成，它阅读UAG密码子，[8]和它的紧密结构类似物3。[9]后一种化合物，由于末端炔基的存在，通过CuI催化的叠氮-炔环加成反应(CuAAC)，使所得到的蛋白质能够与叠氮基荧光团进行位置特异性的翻译后修饰。我们最近的工作已经超越了蛋白质点击化学领域，导致了半胱氨酸衍生物4的合成和应用，用于蛋白质泛素化的天然化学连接。[10]3广泛使用的主要障碍来自于它的难得性和与其合成长度相关的高成本(从抗坏血酸到16步)。[9]2和3设计的指导原则是希望保持1和其类似物之间的空间和电子相似性，因为这被认为是成功的吡咯赖氨酸模拟的先决条件。Polycarpo等人[1]早些时候关于简单的吡咯赖氨酸类似物的报告似乎强化了这一观点。特别是，据报道，在N-6位含有非环酰基的赖氨酸衍生物不能阅读UAG密码子。然而，Yanagisawa等人最近的一项工作[11]显示，一些非环类似物也可以作为可行的吡咯赖氨酸替代品。此外，据报道，即使是N6-乙酰赖氨酸，当提供高浓度的[12]或在Pyls(吡咯酰-tRNA合成酶)突变体存在的情况下，[13]也可以带电到PylT(TRNAPyl)上，甚至结合到重组蛋白中。
The site-specific tagging of proteins with organic fluorophores [2] has been proven to be a powerful method in structural and functional studies of a wide range of biological systems.[3-5] The most challenging aspect of this approach is the regiospecific incorporation of a suitable fluorophore on or nearby a rationally selected amino acid residue within a protein chain. The demonstration that pyrrolysine (1, Figure 1), the 22nd geneticallyencoded amino acid, can be incorporated into recombinant proteins in response to the UAG codon,[6, 7] prompted us to search for similarly incorporable analogs of 1 harboring reactive functionalities suitable for anchoring small organic fluorophores. We previously reported the synthesis of the THF-containing lysine derivative 2 that reads through the UAG codon,[8] as does its close structural analog 3.[9] The latter compound, thanks to the presence of the terminal alkyne functionality, enables site-specific post-translational modification of the resulting protein with azide-based fluorophores via the CuI-catalyzed azide–alkyne cycloaddition reaction (CuAAC). Our recent work has gone beyond the field of protein click chemistry resulting in the synthesis and application of the cysteine derivative 4 for protein ubiquitination via native chemical ligation.[10]The main obstacle in the widespread use of 3 stems from its inaccessibility and high cost associated with the length of its synthesis (16 steps from ascorbic acid).[9] The guiding principle in the design of both 2 and 3 was the desire to maintain the steric and electronic similarity between 1 and its analogues, as it was believed to be a prerequisite for a successful pyrrolysine mimic. An earlier report by Polycarpo et al.[1] on simple pyrrolysine analogs had seemed to reinforce this notion. In particular, lysine derivatives bearing an acyclic acyl substituent at N-6 had been reported not to read through the UAG codon. However, a more recent work by Yanagisawa et al.[11] revealed that some acyclic analogs can also serve as viable pyrrolysine substitutes. Moreover, it was also reported that even N6-acetyllysine, when supplied at high concentration [12] or in the presence of a PylS (pyrrolysyl-tRNA synthetase) mutant,[13] can be charged onto PylT (tRNAPyl) or even incorporated into recombinant proteins.
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影响因子：
56.9
作者：
IKURA, M;CLORE, GM;BAX, A
通讯作者：
BAX, A
影响因子：
--
作者：
Yanagisawa, Tatsuo;Ishii, Ryohei;Yokoyama, Shigeyuki
通讯作者：
Yokoyama, Shigeyuki
影响因子：
64.8
作者：
van Kasteren, Sander I.;Kramer, Holger B.;Davis, Benjamin G.
通讯作者：
Davis, Benjamin G.
影响因子：
48
作者：
Liu, Wenshe;Brock, Ansgar;Schultz, Peter G.
通讯作者：
Schultz, Peter G.
影响因子：
--
作者：
JONES, G;JACKSON, WR;BERGMARK, WR
通讯作者：
BERGMARK, WR