Detection of Alkynes via Click Chemistry with a Brominated Coumarin Azide by Simultaneous Fluorescence and Isotopic Signatures in Mass Spectrometry.
Detection of Alkynes via Click Chemistry with a Brominated Coumarin Azide by Simultaneous Fluorescence and Isotopic Signatures in Mass Spectrometry.
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通过质谱中同时荧光和同位素特征，利用溴化香豆素叠氮化物进行点击化学检测炔烃。
DOI：
10.1021/acs.bioconjchem.7b00354
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发表时间：
2017
影响因子：
4.7
通讯作者：
Zhou,ZhaohuiSunny
中科院分区：
文献类型：
--
作者：
Yang,Lihua;Chumsae,Chris;Kaplan,JeniferB;Moulton,KevinRyan;Wang,Dongdong;Lee,DavidH;Zhou,ZhaohuiSunny
炔是点击化学的关键组分，并且用于各种各样的应用，包括生物缀合、蛋白质修饰的选择性标记以及代谢物和药物靶标的标记。然而，对于检测炔仍然存在挑战，因为来自炔和叠氮化物的大多数1，2，3-三唑产物不具有可用于通过荧光或质谱法对其进行简易检测的独特固有性质。为了解决这一关键需求，一种新的溴化香豆素叠氮化物被用来标记炔和检测炔共轭的生物分子。该标签具有几个有用的特性：第一，它是荧光的，点击化学产物是高度荧光和可量化的;第二，其独特的同位素模式有助于通过质谱法进行识别;第三，其点击化学产物在片段化后形成独特的报告离子对，可用于快速筛选数据。使用与炔缀合的单克隆抗体，已经开发了一般工作流程并进行了全面检查。
Alkynes are a key component of click chemistry and used for a wide variety of applications including bioconjugation, selective tagging of protein modifications, and labeling of metabolites and drug targets. However, challenges still exist for detecting alkynes because most 1,2,3-triazole products from alkynes and azides do not possess distinct intrinsic properties that can be used for their facile detection by either fluorescence or mass spectrometry. To address this critical need, a novel brominated coumarin azide was used to tag alkynes and detect alkyne-conjugated biomolecules. This tag has several useful properties: first, it is fluorogenic and the click-chemistry products are highly fluorescent and quantifiable; second, its distinct isotopic pattern facilitates identification by mass spectrometry; and third, its click-chemistry products form a unique pair of reporter ions upon fragmentation that can be used for the quick screening of data. Using a monoclonal antibody conjugated with alkynes, a general workflow has been developed and examined comprehensively.