Highly efficient and chemoselective blocking of free amino group by ortho-phthalaldehyde (OPA) for comprehensive analysis of protein terminome

Ting Wang; Zhongjie Wang; Rui Wang; Lei Zhang; Ying Zhang; Haojie Lu

doi:10.1016/j.talanta.2023.125262

Highly efficient and chemoselective blocking of free amino group by ortho-phthalaldehyde (OPA) for comprehensive analysis of protein terminome

Talanta. 2024 Jan 15:267:125262. doi: 10.1016/j.talanta.2023.125262. Epub 2023 Oct 4.

Authors

Ting Wang¹, Zhongjie Wang², Rui Wang², Lei Zhang², Ying Zhang³, Haojie Lu⁴

Affiliations

¹ Department of Chemistry and Liver Cancer Institute of Zhongshan Hospital, Fudan University, Shanghai, 200032, China.
² Institutes of Biomedical Sciences and NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai, 200032, China.
³ Department of Chemistry and Liver Cancer Institute of Zhongshan Hospital, Fudan University, Shanghai, 200032, China; Institutes of Biomedical Sciences and NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai, 200032, China. Electronic address: ying@fudan.edu.cn.
⁴ Department of Chemistry and Liver Cancer Institute of Zhongshan Hospital, Fudan University, Shanghai, 200032, China; Institutes of Biomedical Sciences and NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai, 200032, China. Electronic address: luhaojie@fudan.edu.cn.

PMID: 37804787
DOI: 10.1016/j.talanta.2023.125262

Abstract

Herein, we introduced ortho-phthalaldehyde (OPA) for blocking free amino groups and established a simple and robust method for comprehensive profiling of protein terminome based on strong cation exchange chromatography (SCX) fractionation. With the highly efficient and chemoseletive amine-group blocking, we identified 2271 canonical human protein N-termini, 1650 canonical human protein C-termini, as well as 645 protein neo-N-termini from HeLa cells.

Keywords: Comprehensive analysis; Neo-N-Termini; OPA; Protein terminome.

MeSH terms

HeLa Cells
Humans
Proteins*
o-Phthalaldehyde* / chemistry

Substances

o-Phthalaldehyde
Proteins