Dichloromaleimide (diCMI): A Small and Fluorogenic Reactive Group for Use in Affinity Labeling
Dichloromaleimide (diCMI): A Small and Fluorogenic Reactive Group for Use in Affinity Labeling
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DOI：
10.1248/cpb.c16-00501
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发表时间：
2016-11-01
影响因子：
1.7
通讯作者：
Yamaguchi, Takao
中科院分区：
文献类型：
--
作者：
Chiba, Kosuke;Hashimoto, Yuichi;Yamaguchi, Takao
包含配体部分、反应基团（例如环氧化物、卤代乙酰基或光反应基团）和标签单元（例如荧光团或放射性同位素）的化学探针广泛用于亲和标记以识别生物活性分子的靶蛋白。然而，高功能化化学探针的设计和合成通常非常耗时。在本文中，我们提出了一种带有小2,3-二氯马来酰亚胺（diCMI）单元的化学探针的简单设计策略，该单元通过与目标蛋白配体结合位点附近的亲核赖氨酸残基反应生成2-氨基-3-氯马来酰亚胺荧光团，作为组合反应基团和标签单元。模型配体蛋白实验证实 diCMI 单元具有合适的反应性和荧光能力，可进行有效的亲和标记。
Chemical probes comprising a ligand moiety, a reactive group (e.g. epoxide, haloacetyl or photoreactive group) and a tag unit (e.g. fluorophore or radioisotope) are widely used in affinity labeling to identify the target proteins of bioactive molecules. However, design and synthesis of highly functionalized chemical probes are often time-consuming. In this paper, we propose a simple design strategy for chemical probes bearing a small 2,3-dichloromaleimide (diCMI) unit, which serves as a combined reactive group and tag unit by reacting with a nucleophilic lysine residue near the ligand-binding site of the target protein to generate the 2-amino-3-chloromaleimide fluorophore. Model ligand protein experiments confirmed that the diCMI unit has suitable reactivity and fluorogenic capability for efficient affinity labeling.