通过耦合稳定同位素示踪和诱导电喷雾质谱法对活细胞中的代谢物进行谱图分析和鉴定。

Metabolite profiling and identification in living cells by coupling stable isotope tracing and induced electrospray mass spectrometry.

作者信息

Hou Zhuanghao, Zhan Liujuan, Cao Kaiming, Luan Moujun, Wang Xinchen, Zhang Buchun, Ma Likun, Yin Hao, Liu Zhicheng, Liu Yangzhong, Huang Guangming

机构信息

Department of Cardiology, The First Affiliated Hospital of USTC, University of Science and Technology of China, 230001, Hefei, China; School of Chemistry and Materials Science, University of Science and Technology of China, 230026, Hefei, China.

出版信息

Anal Chim Acta. 2023 Feb 8;1241:340795. doi: 10.1016/j.aca.2023.340795. Epub 2023 Jan 4.

DOI:10.1016/j.aca.2023.340795

PMID:36657872

Abstract

Direct observation of metabolites in living cells by mass spectrometry offers a bright future for biological studies but also suffers a severe challenge to untargeted peak assignment to tentative metabolite candidates. In this study, we developed a method combining stable isotope tracing and induced electrospray mass spectrometry for living-cells metabolite measurement and identification. By using C-glucose and ammonium chloride-N as the sole carbon and nitrogen sources for cell culture, Escherichia coli synthesized metabolites with N and C elements. Tracing the number of carbon and nitrogen atoms could offer a complementary dimension for candidate peak searching. As a result, the identification confidence of metabolites achieved a universal improvement based on carbon/nitrogen labelling and filtration.

摘要

通过质谱直接观察活细胞中的代谢物为生物学研究带来了光明的前景，但在将未靶向的峰分配给暂定代谢物候选物方面也面临严峻挑战。在本研究中，我们开发了一种结合稳定同位素示踪和诱导电喷雾质谱的方法，用于活细胞代谢物的测量和鉴定。通过使用¹³C-葡萄糖和¹⁵N-氯化铵作为细胞培养的唯一碳源和氮源，大肠杆菌合成了含有¹³C和¹⁵N元素的代谢物。追踪碳和氮原子的数量可为候选峰搜索提供一个补充维度。结果，基于碳/氮标记和过滤，代谢物的鉴定可信度得到了普遍提高。