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Researchers Tailor Rapid and Effective N-phosphopeptides Enrichment Strategy

Jan 11, 2021

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) proposed a rapid and effective N-phosphopeptides enrichment strategy.

Their study was published in Nature Communications on Dec. 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 provide 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, 3384 N-pho sites, containing 611 pHis, 1618 pLys and 1155 pArg, were identified from HeLa cell lysates.

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

This study was supported by 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.

Contact

WANG Yongjin

Dalian Institute of Chemical Physics

E-mail:

Bis(zinc(II)-dipicolylamine)-functionalized sub-2 μm core-shell microspheres for the analysis of N-phosphoproteome

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