Zhou Xiaoyu, Zhang Hongyan, Wang Li, Lv Liting, Wu Ren'an
Laboratory of High-Resolution Mass Spectrometry Technologies, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China.
CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China.
Analyst. 2023 Mar 27;148(7):1483-1491. doi: 10.1039/d2an02004a.
Protein glycosylation and phosphorylation play essential roles in biological systems. The crosstalk of both glycosylation and phosphorylation on one protein represents an unveiled biological function. To realize the analyses of both glycopeptides and phosphopeptides, a simultaneous enrichment method of -glycopeptides, mono-phosphopeptides and multi-phosphopeptides was developed based on a multi-functional dual-metal centered zirconium metal-organic framework that provided multiple interactions for HILIC, IMAC, and MOAC for glycopeptides and phosphopeptides. Based on a careful optimization of sample loading and elution conditions for the simultaneous enrichment of glycopeptides and phosphopeptides with the zirconium metal-organic framework, a total of 1011 -glycopeptides derived from 410 glycoproteins and 1996 phosphopeptides including 741 multi-phosphopeptides derived from 1189 phosphoproteins could be identified from a HeLa cell digest. The simultaneous enrichment approach for glycopeptides and mono-/multi-phosphopeptides demonstrates the great potential of the combined interactions for HILIC, IMAC, and MOAC in integrated post-translational modification proteomics research.
蛋白质糖基化和磷酸化在生物系统中发挥着重要作用。同一蛋白质上糖基化和磷酸化的相互作用代表了一种尚未揭示的生物学功能。为了实现对糖肽和磷酸肽的分析，基于一种多功能双金属中心锆金属有机框架开发了一种同时富集O-糖肽、单磷酸肽和多磷酸肽的方法，该框架为糖肽和磷酸肽的亲水作用色谱（HILIC）、固定金属离子亲和色谱（IMAC）和金属氧化物亲和色谱（MOAC）提供了多种相互作用。基于对锆金属有机框架同时富集糖肽和磷酸肽的样品加载和洗脱条件的仔细优化，从HeLa细胞消化物中可以鉴定出总共1011个来自410种糖蛋白的O-糖肽和1996个磷酸肽，其中包括741个来自1189种磷蛋白的多磷酸肽。糖肽和单/多磷酸肽的同时富集方法证明了HILIC、IMAC和MOAC的联合相互作用在整合的翻译后修饰蛋白质组学研究中的巨大潜力。
