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Researchers Tailor Rapid and Effective N-phosphopeptides Enrichment Strategy
  English.dicp.cas.cn    Posted:2021-01-11    From:Group 1810

Protein N-phosphorylation plays a critical role in central metabolism and two/multicomponent signaling of prokaryotes. Furthermore, it exists as intermediates of some important enzymes in mammals.

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.

Recently, a research group led by Prof. ZHANG Lihua and Pro. ZHANG Yukui from the Dalian Institute of Chemical Physics (DICP) of the Chinese Academy of Sciences (CAS) reported a rapid and effective N-phosphopeptides enrichment strategy.

This results were published in Nature Communications on December 4.


Bis(zinc(II)-dipicolylamine)-functionalized sub-2 μm core-shell microspheres for the analysis of N-phosphoproteome (Image by JIANG Bo and HU Yechen) 

Inspired by naturally occurring phosphatases, which provided specific Zn(II)-central enzymatic pockets to bind phosphate units of substrates, the researchers designed bis(zinc(II)-dipicolylamine) molecular (DpaZn) for phosphate targets recognition under neutral conditions.

Combing the fast mass transfer ability of sub-2 μm core-shell silica microspheres, DpaZn-functionalized sub-2 μm core-shell silica microspheres (SiO2@DpaZn) were designed for on-tip N-phosphopeptides enrichment under neutral conditions. In total, 3,384 N-pho sites, containing 611 pHis, 1,618 pLys and 1,155 pArg, were identified from HeLa cell lysates.

The new N-phosphorylated proteome analysis method not only provides basic data for in-depth study of its biological function, but also provides technical support for promoting the development of precision medicine, synthetic biology and other fields.

This study was supported by the foundations and key foundation from the National Natural Science Foundation of China, the National Key Research and Development Program of China, and the innovation program of science and research from DICP, CAS. (Text by JIANG Bo and HU Yechen)

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