Beijing National Laboratory for Molecular Sciences, National Centre for Mass Spectrometry in Beijing, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, People's Republic of China.
College of Chemistry and Materials Science, Key Laboratory of Functional Molecular Solids, the Ministry of Education, Anhui Laboratory of Molecular-Based Materials, Anhui Normal University, Wuhu, Anhui 241000, People's Republic of China.
Anal Chem. 2020 Dec 1;92(23):15517-15525. doi: 10.1021/acs.analchem.0c03448. Epub 2020 Nov 17.
visualization of proteins of interest in single cells is attractive in cell biology, molecular biology, and biomedicine fields. Time-of-flight-secondary ion mass spectrometry (ToF-SIMS) is a powerful tool for imaging small organic molecules in single cells, yet difficult to image biomacromolecules such as proteins and DNA. Herein, a universal strategy is reported to image specific proteins in single cells by ToF-SIMS following genetic incorporation of fluorine-containing unnatural amino acids as a chemical tag into the proteins a genetic code expansion technique. The method was developed and validated by imaging a green fluorescence protein (GFP) in and human HeLa cancer cells and then utilized to visualize the characteristic polar distribution of chemotaxis protein CheA in cells and the interaction between high-mobility group box 1 protein and cisplatin-damaged DNA in HeLa cells. The present work highlights the power of ToF-SIMS imaging combined with genetically encoded chemical tags for visualization of specific proteins as well as the interactions between proteins and drugs or drug-damaged DNA in single cells.
在细胞生物学、分子生物学和生物医学领域，对单细胞中感兴趣的蛋白质进行可视化是很有吸引力的。飞行时间二次离子质谱（ToF-SIMS）是一种对单细胞中小分子有机分子成像的强大工具，但很难对蛋白质和 DNA 等生物大分子进行成像。在此，报道了一种通过将含氟非天然氨基酸作为化学标记物通过遗传密码扩展技术掺入蛋白质中，然后用 ToF-SIMS 对单细胞中特定蛋白质进行成像的通用策略。该方法通过对 和人宫颈癌细胞中的绿色荧光蛋白（GFP）进行成像来开发和验证，然后用于可视化趋化蛋白 CheA 在 细胞中的特征极性分布以及高迁移率族框 1 蛋白与顺铂损伤 DNA 在 HeLa 细胞中的相互作用。本工作突出了将遗传编码化学标记物与 ToF-SIMS 成像相结合用于可视化单细胞中特定蛋白质以及蛋白质与药物或药物损伤 DNA 之间相互作用的强大功能。
