Issue 22, 2019

Capillary zone electrophoresis-tandem mass spectrometry with ultraviolet photodissociation (213 nm) for large-scale top–down proteomics

Abstract

Capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) has attracted attention recently for large-scale top–down proteomics that aims to characterize proteoforms in cells at a global scale with high throughput. In this work, CZE-MS/MS with ultraviolet photodissociation (UVPD) was evaluated for large-scale top–down proteomics for the first time. Roughly, 600 proteoforms and 369 proteins were identified from a zebrafish brain sample via coupling size exclusion chromatography (SEC) fractionation with CZE-UVPD. The dataset represents one of the largest top–down proteomics datasets using UVPD. Single-shot CZE-UVPD identified 227 proteoforms of 139 proteins from one SEC fraction of the zebrafish brain sample. The SEC-CZE-UVPD system identified zebrafish brain proteoforms in a mass range of 3–21 kDa. UVPD with 213 nm photons produced reasonably good gas-phase fragmentation of proteoforms. For instance, 75% backbone cleavage was observed for parvalbumin-7 with about 12 kDa molecular weight. The system detected various post-translational modifications (PTMs) from the zebrafish brain sample, including N-terminal acetylation, trimethylation and myristoylation of N-terminal glycine. Two different proteoforms of calmodulin, with either only N-terminal acetylation or both N-terminal acetylation and K115 trimethylation, were identified in the zebrafish brain sample. To the best of our knowledge, there is no experimental evidence reported in the literature on the two proteoforms of calmodulin in zebrafish brain.

Graphical abstract: Capillary zone electrophoresis-tandem mass spectrometry with ultraviolet photodissociation (213 nm) for large-scale top–down proteomics

Supplementary files

Article information

Article type
Communication
Submitted
21 mars 2019
Accepted
05 mai 2019
First published
07 mai 2019

Anal. Methods, 2019,11, 2855-2861

Capillary zone electrophoresis-tandem mass spectrometry with ultraviolet photodissociation (213 nm) for large-scale top–down proteomics

E. N. McCool, D. Chen, W. Li, Y. Liu and L. Sun, Anal. Methods, 2019, 11, 2855 DOI: 10.1039/C9AY00585D

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements