Native Proteomics by Capillary Zone Electrophoresis-Mass Spectrometry

Overview

Journal Angew Chem Int Ed Engl

Specialty Chemistry

Date 2024 Aug 28

PMID 39196601

Authors

Qianyi Wang

Qianjie Wang

Zihao Qi

William Moeller

Vicki H Wysocki

Liangliang Sun

Affiliations

Soon will be listed here.

Abstract

Native proteomics measures endogenous proteoforms and protein complexes under a near physiological condition using native mass spectrometry (nMS) coupled with liquid-phase separations. Native proteomics should provide the most accurate bird's-eye view of proteome dynamics within cells, which is fundamental for understanding almost all biological processes. nMS has been widely employed to characterize well-purified protein complexes. However, there are only very few trials of utilizing nMS to measure proteoforms and protein complexes in a complex sample (i.e., a whole cell lysate). Here, we pioneer the native proteomics measurement of large proteoforms or protein complexes up to 400 kDa from a complex proteome via online coupling of native capillary zone electrophoresis (nCZE) to an ultra-high mass range (UHMR) Orbitrap mass spectrometer. The nCZE-MS technique enabled the measurement of a 115-kDa standard protein complex while consuming only about 0.1 ng of protein material. nCZE-MS analysis of an E.coli cell lysate detected 72 proteoforms or protein complexes in a mass range of 30-400 kDa in a single run while consuming only 50-ng protein material. The mass distribution of detected proteoforms or protein complexes agreed well with that from mass photometry measurement. This work represents a technical breakthrough in native proteomics for measuring complex proteomes.

Citing Articles

Mass Spectrometry-Based Top-Down Proteomics in Nanomedicine: Proteoform-Specific Measurement of Protein Corona.

Sadeghi S, Ashkarran A, Wang Q, Zhu G, Mahmoudi M, Sun L ACS Nano. 2024; .

PMID: 39276099 PMC: 11440641. DOI: 10.1021/acsnano.4c04675.

The role of protein corona in advancing plasma proteomics.

Saei A, Sun L, Mahmoudi M Proteomics. 2024; 25(1-2):e2400028.

PMID: 39221533 PMC: 11735278. DOI: 10.1002/pmic.202400028.

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