Proteomics

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


ABSTRACT: Herein, bis(zinc(II)-dipicolylamine) functionalized sub-2 μm core-shell silica microspheres (SiO2@DpaZn) were tailored for N-phosphopeptide enrichment at pH 7.7. Contributed by the coordination bonding between Zn(II) and phosphate groups with the KD value of 13.14 μM, N-phosphopeptides could be captured efficiently under neutral conditions. Moreover, contributed by the core-shell structure of SiO2@DpaZn, the fast on-tip enrichment ensured the recovery of N-phosphopeptides up to 44.1%, 3-fold higher than that obtained by in-solution capture. Moreover, 40 determined N-pho sites on 32 substrates were identified in E. coli lysates, among which five had kinase activity. All these results demonstrate that our method is beneficial to promote the discovery of N-phosphoproteins with significant biological functions.

ORGANISM(S): Homo Sapiens

SUBMITTER: Lihua Zhang  

PROVIDER: PXD021067 | iProX | Sat Aug 22 00:00:00 BST 2020

REPOSITORIES: iProX

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

Hu Yechen Y   Jiang Bo B   Weng Yejing Y   Sui Zhigang Z   Zhao Baofeng B   Chen Yuanbo Y   Liu Lukuan L   Wu Qiong Q   Liang Zhen Z   Zhang Lihua L   Zhang Yukui Y  

Nature communications 20201204 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<sub>2</sub>@DpaZn) are tailored for rapid  ...[more]

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