Liu Zhen, Liang Yong, Zhou Yang, Ge Fuchun, Yan Xiaowen, Yang Limin, Wang Qiuquan
Department of Chemistry & the MOE Key Lab of Spectrochemical Analysis and Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
iScience. 2021 Apr 3;24(5):102397. doi: 10.1016/j.isci.2021.102397. eCollection 2021 May 21.
Fucosylation and its fucosidic linkage-specific motifs are believed to be essential to understand their distinct roles in cellular behavior, but their quantitative information has not yet been fully disclosed due to the requirements of ultra-sensitivity and selectivity. Herein, we report an approach that converts fucose (Fuc) to stable europium (Eu) isotopic mass signal on hard ionization inductively coupled plasma mass spectrometry (ICP-MS). Metabolically assembled -fucose on the cell surface allows us to tag them with an -customized Eu-crafted bacteriophage MS2 capsid nanoparticle for Eu signal multiplication, resulting in an ever lowest detection limit of 4.2 mol Fuc. Quantitative breakdown of the linkage-specific fucosylation motifs in situ preserved on single cancerous HepG2 and paracancerous HL7702 cells can thus be realized on a single-cell ICP-MS platform, specifying their roles during the cancering process. This approach was further applied to the discrimination of normal hepatocellular cells and highly, moderately, and poorly differentiated hepatoma cells collected from real hepatocellular carcinoma tissues.
岩藻糖基化及其岩藻糖苷键特异性基序被认为对于理解它们在细胞行为中的独特作用至关重要,但由于对超灵敏度和选择性的要求,其定量信息尚未完全揭示。在此,我们报告了一种方法,该方法可在硬电离电感耦合等离子体质谱(ICP-MS)上将岩藻糖(Fuc)转化为稳定的铕(Eu)同位素质量信号。细胞表面代谢组装的α-岩藻糖使我们能够用定制的铕修饰的噬菌体MS2衣壳纳米颗粒对其进行标记,以实现Eu信号倍增,从而实现了低至4.2 nmol Fuc的检测限。因此,可以在单细胞ICP-MS平台上实现对原位保存在单个肝癌HepG2细胞和癌旁HL7702细胞上的连接特异性岩藻糖基化基序的定量分析,明确它们在癌变过程中的作用。该方法进一步应用于从实际肝癌组织中收集的正常肝细胞与高分化、中分化和低分化肝癌细胞的鉴别。