Hu Yechen, Jiang Bo, Weng Yejing, Sui Zhigang, Zhao Baofeng, Chen Yuanbo, Liu Lukuan, Wu Qiong, Liang Zhen, Zhang Lihua, Zhang Yukui
CAS Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R & A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 116023, Dalian, China.
University of Chinese Academy of Sciences, 100049, Beijing, China.
Nat Commun. 2020 Dec 4;11(1):6226. doi: 10.1038/s41467-020-20026-1.
Protein N-phosphorylation plays a critical role in central metabolism and two/multicomponent signaling of prokaryotes. However, the current enrichment methods for O-phosphopeptides are not preferred for N-phosphopeptides due to the intrinsic lability of P-N bond under acidic conditions. Therefore, the effective N-phosphoproteome analysis remains challenging. Herein, bis(zinc(II)-dipicolylamine)-functionalized sub-2 μm core-shell silica microspheres (SiO@DpaZn) are tailored for rapid and effective N-phosphopeptides enrichment. Due to the coordination of phosphate groups to Zn(II), N-phosphopeptides can be effectively captured under neutral conditions. Moreover, the method is successfully applied to an E.coli and HeLa N-phosphoproteome study. These results further broaden the range of methods for the discovery of N-phosphoproteins with significant biological functions.
蛋白质N-磷酸化在原核生物的中心代谢和二元/多组分信号传导中起着关键作用。然而，由于P-N键在酸性条件下具有内在不稳定性，目前用于O-磷酸肽的富集方法并不适用于N-磷酸肽。因此，有效的N-磷酸蛋白质组分析仍然具有挑战性。在此，双（锌（II）-二吡啶甲胺）功能化的亚2μm核壳二氧化硅微球（SiO@DpaZn）被定制用于快速有效地富集N-磷酸肽。由于磷酸基团与Zn（II）的配位作用，N-磷酸肽可以在中性条件下被有效捕获。此外，该方法已成功应用于大肠杆菌和HeLa细胞的N-磷酸蛋白质组研究。这些结果进一步拓宽了发现具有重要生物学功能的N-磷酸化蛋白质的方法范围。
