Abstract: A coumarin-based probe coumarin-alkene ketone-benzylaminoguanine(CAKBG)containing both α, β-unsaturated ketone and SNAP-tag^TM substrate was synthesized. By testing the UV-Vis absorption spectra and fluorescence emission spectra of the probe CAKBG, the results show that the probe CAKBG has a longer absorption wavelength in polar solvents( such as glycerol, ethylene glycol, water, DMSO), and the probe CAKBG is not sensitive to the pH change of the solutions. In addition, spectral experiments show that both Cys and Hcy can cause a blue shift in the maximum absorption wavelength of the probe CAKBG, and when the excitation wavelength is 405 nm, both Cys and Hcy can cause the fluorescence emission peak of the probe CAKBG at 487 nm to be significantly enhanced. Cell imaging results show that the thiol fluorescent probe CAKBG containing SNAP-tag^TM substrate can image Cys and Hcy in living cells. The above results indicate that SNAP-Tag^TM protein labeling technology is expected to be widely- DOI:10.3969/j.issn.1000-1565.2024.05.007一种含SNAP-tag^TM底物的硫醇荧光探针林利霞¹,郭春青¹,王利斌¹,赵子彪¹,熊康明^2,3(1.山西警察学院 治安系,山西 太原 030401;2.山西白求恩医院 纳米成像与载药制剂山西省重点实验室,山西 太原 030032;3.中国科学院大连化学物理研究所 分离分析化学重点实验室,辽宁 大连 116023)摘 要:合成了一种同时含α, β-不饱和酮活性位点和SNAP-tag^TM底物的香豆素母体探针香豆素-烯酮-苯甲氨基鸟嘌呤(CAKBG).通过测试探针CAKBG的紫外-可见吸收光谱和荧光发射光谱,发现探针CAKBG在极性溶剂(例如, 甘油、乙二醇、水、DMSO)中有更长的吸收波长,且探针CAKBG对溶液的pH变化不敏感.此外,光谱实验发现生物硫醇Cys和Hcy均能引起探针CAKBG的最大吸收波长发生蓝移,且当激发波长为405 nm时,Cys和Hcy均能导致探针CAKBG在487 nm处的荧光发射峰明显增强.细胞成像结果表明,含有SNAP-tag^TM底物的硫醇荧光探针CAKBG在活细胞中可以对Cys、Hcy进行荧光成像.综上,SNAP-Tag^TM蛋白标签技术有望今后在硫醇荧光探针的细胞成像中得到推广应用.关键词:α, β-不饱和酮;SNAP-Tag^TM底物;硫醇荧光探针;细胞成像中图分类号:O622.4 文献标志码:A 文章编号:1000-1565(2024)05-0504-09A thiol fluorescent probe containing SNAP-tag^TM substrateLIN Lixia¹, GUO Chunqing¹, WANG Libin¹, ZHAO Zibiao¹, XIONG Kangming^2,3(1. Department of Security, Shanxi Police College, Taiyuan 030401, China; 2. Key Laboratory of Nano-imaging and Drugloaded Preparation of Shanxi Province, Shanxi Bethune Hospital, Taiyuan 030032, China; 3. CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China)Abstract: A coumarin-based probe coumarin-alkene ketone-benzylaminoguanine(CAKBG)containing both α, β-unsaturated ketone and SNAP-tag^TM substrate was synthesized. By testing the UV-Vis absorption spectra and fluorescence emission spectra of the probe CAKBG, the results show that the probe CAKBG has a longer absorption wavelength in polar solvents( such as glycerol, ethylene glycol, water, DMSO), and the probe CAKBG is not sensitive to the pH change of the solutions. In addition, spectral experiments show that both Cys and Hcy can cause a blue shift in the maximum absorption wavelength of the probe CAKBG, and when the excitation wavelength is 405 nm, both Cys and Hcy can cause the fluorescence emission peak of the probe CAKBG at 487 nm to be significantly enhanced. Cell imaging results show that the thiol fluorescent probe CAKBG containing SNAP-tag^TM substrate can image Cys and Hcy in living cells. The above results indicate that SNAP-Tag^TM protein labeling technology is expected to be widely- 收稿日期:2023-06-15;修回日期:2023-09-12 基金项目:国家自然科学基金资助项目(22008232);山西省基础研究计划青年科学研究项目(202203021212512) 第一作者:林利霞(1991—),女,山西警察学院讲师,博士,主要从事有机小分子的光谱性能研究. E-mail:1552774577@qq.com 通信作者:熊康明(1990—),男,山西白求恩医院副研究员,博士,主要从事荧光探针的传感识别研究. E-mail:1187994460@qq.com 第5期林利霞等:一种含SNAP-tag^TM底物的硫醇荧光探针河北大学学报(自然科学版) 第44卷used in cell imaging of thiol fluorescent probes in the future.

Key words: α, β-unsaturated ketone, SNAP-Tag^TM substrate, thiol fluorescent probes, cell imaging