Discovery of a Fluorogenic Probe for In Situ Pyruvate Kinase M2 Isoform (PKM2) Labeling through Chemoselective SNAr with a Binding Site Lysine Residue
Discovery of a Fluorogenic Probe for In Situ Pyruvate Kinase M2 Isoform (PKM2) Labeling through Chemoselective SNAr with a Binding Site Lysine Residue
复制标题
通过具有结合位点赖氨酸残基的化学选择性 SNAr 发现用于原位丙酮酸激酶 M2 同工型 (PKM2) 标记的荧光探针
DOI：
10.1021/acs.analchem.1c00208
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发表时间：
2021
影响因子：
7.4
通讯作者：
Zhang Zhichao
中科院分区：
文献类型：
--
作者：
Wang Ziqian;Zhang Xiaodong;Zhang Hong;Tang Yao;Pan Hao;Wang Hang;Ji Tong;Guo Yafei;Gao Qishuang;Song Ting;Zhang Zhichao
开发基于反应的开启探针的关键挑战是建立仅在与特定蛋白质结合时表现出高反应性的潜在亲电子荧光团。在此，我们鉴定了这样一种荧光团，即 6-芳基硫醚取代的 3-cyano-1-oxo-1H-phenalene-2-carboxylate，它化学选择性地标记结合位点 Cys 或 Lys 残基。基于该荧光团，我们开发了第一个基于反应的丙酮酸激酶M2亚型（PKM2）荧光探针AT-OPC1，它用结合位点Lys305选择性地标记PKM2。荧光团的潜在亲电反应性使探针能够通过蛋白质组水平的凝胶内荧光成像和实时免洗细胞成像方法原位精确检测PKM2的表达，具有在癌症诊断中应用的潜力。
The key challenge of developing reaction-based turn-on probes is to establish latent electrophilic fluorophores exhibiting high reactivity only upon binding to a specific protein(s). Herein, we identified such a fluorophore, 6-arylthioether-substituted 3-cyano-1-oxo-1H-phenalene-2-carboxylate, which chemoselectively labels binding site Cys or Lys residues. Based on this fluorophore, we developed the first reaction-based turn-on pyruvate kinase M2 isoform (PKM2) fluorescent probeAT-OPC1, which selectively labels PKM2 with the binding site Lys305. The latent electrophilic reactivity of the fluorophore endows the probe with precise detection of the expression of PKM2 in situ by means of both in-gel fluorescence imaging at the proteome level and real-time no-wash cell imaging approaches, which has the potential to be applied in cancer diagnoses.