Transglutaminase-Catalyzed Bioconjugation Using One-Pot Metal-Free Bioorthogonal Chemistry,Bioconjugate Chemistry

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Transglutaminase-Catalyzed Bioconjugation Using One-Pot Metal-Free Bioorthogonal Chemistry
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2017-09-14 00:00:00 , DOI: 10.1021/acs.bioconjchem.7b00509
Natalie M. Rachel _{1,

2} , Jacynthe L. Toulouse ₁ , Joelle N. Pelletier _{1,

2}

Affiliation

General approaches for controlled protein modification are increasingly sought-after in the arena of chemical biology. Here, using bioorthogonal reactions, we present combinatorial chemoenzymatic strategies to effectuate protein labeling. A total of three metal-free conjugations were simultaneously or sequentially incorporated in a one-pot format with microbial transglutaminase (MTG) to effectuate protein labeling. MTG offers the particularity of conjugating residues within a protein sequence rather than at its extremities, providing a route to labeling the native protein. The reactions are rapid and circumvent the incompatibility posed by metal catalysts. We identify the tetrazine ligation as most-reactive for this purpose, as demonstrated by the fluorescent labeling of two proteins. The Staudinger ligation and strain-promoted azide–alkyne cycloaddition are alternatives. Owing to the breadth of labels that MTG can use as a substrate, our results demonstrate the versatility of this system, with the researcher being able to combine specific protein substrates with a variety of labels.

中文翻译：

使用一锅无金属的生物正交化学进行转谷氨酰胺酶催化的生物缀合

在化学生物学领域，控制蛋白质修饰的通用方法越来越受欢迎。在这里，使用生物正交反应，我们提出了组合化学酶策略来实现蛋白质标记。将总共三个无金属的缀合物与微生物转谷氨酰胺酶（MTG）以一锅的形式同时或依次掺入以实现蛋白质标记。MTG提供了在蛋白质序列内而不是在其末端缀合残基的特殊性，从而提供了标记天然蛋白质的途径。该反应是快速的并且避免了金属催化剂带来的不相容性。我们确定四嗪连接为此目的最反应，如两个蛋白质的荧光标记所示。施陶丁格结扎法和应变促进的叠氮化物-炔烃环加成法是替代方法。由于MTG可以用作底物的标签种类繁多，我们的结果证明了该系统的多功能性，研究人员能够将特定的蛋白质底物与多种标签结合在一起。

更新日期：2017-09-14

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