Chiba Kosuke, Hashimoto Yuichi, Yamaguchi Takao
Institute of Molecular and Cellular Biosciences, The University of Tokyo.
Chem Pharm Bull (Tokyo). 2016;64(11):1647-1653. doi: 10.1248/cpb.c16-00501.
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.
包含配体部分、反应基团（如环氧化物、卤代乙酰基或光反应基团）和标签单元（如荧光团或放射性同位素）的化学探针被广泛用于亲和标记，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很耗时。在本文中，我们提出了一种带有小的2,3-二氯马来酰亚胺（diCMI）单元的化学探针的简单设计策略，该单元通过与靶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，从而作为组合反应基团和标签单元。模型配体-蛋白质实验证实，diCMI单元具有适合高效亲和标记的反应活性和荧光生成能力。
