Enrichment and separation techniques for large-scale proteomics analysis of the protein post-translational modifications

Junfeng Huang; Fangjun Wang; Mingliang Ye; Hanfa Zou

doi:10.1016/j.chroma.2014.10.107

Enrichment and separation techniques for large-scale proteomics analysis of the protein post-translational modifications

J Chromatogr A. 2014 Dec 12:1372C:1-17. doi: 10.1016/j.chroma.2014.10.107. Epub 2014 Nov 6.

Authors

Junfeng Huang¹, Fangjun Wang², Mingliang Ye³, Hanfa Zou⁴

Affiliations

¹ CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China; University of Chinese Academy of Sciences, Beijing, China.
² CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
³ CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China. Electronic address: mingliang@dicp.ac.cn.
⁴ CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China. Electronic address: hanfazou@dicp.ac.cn.

PMID: 25465002
DOI: 10.1016/j.chroma.2014.10.107

Abstract

Comprehensive analysis of the post-translational modifications (PTMs) on proteins at proteome level is crucial to elucidate the regulatory mechanisms of various biological processes. In the past decades, thanks to the development of specific PTM enrichment techniques and efficient multidimensional liquid chromatography (LC) separation strategy, the identification of protein PTMs have made tremendous progress. A huge number of modification sites for some major protein PTMs have been identified by proteomics analysis. In this review, we first introduced the recent progresses of PTM enrichment methods for the analysis of several major PTMs including phosphorylation, glycosylation, ubiquitination, acetylation, methylation, and oxidation/reduction status. We then briefly summarized the challenges for PTM enrichment. Finally, we introduced the fractionation and separation techniques for efficient separation of PTM peptides in large-scale PTM analysis.

Keywords: Enrichment; Mass spectrometry; Multi-dimensional liquid chromatography separation; Post-translational modifications; Protein glycosylation; Protein phosphorylation.

Publication types

Review