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Maleic Anhydride Labeling-Based Approach for Quantitative Proteomics and Successive Derivatization of Peptides

Tian, Shanshan, Zheng, Shuzhen, Han, Yanpu, Guo, Zhenchang, Zhai, Guijin, Bai, Xue, He, Xiwen, Fan, Enguo, Zhang, Yukui, Zhang, Kai
Analytical chemistry 2017 v.89 no.16 pp. 8259-8265
derivatization, hydrophilicity, maleic anhydrides, peptides, proteins, proteomics, stable isotopes
Chemical derivatization is a simple approach for stable-isotope covalent labeling of proteins in quantitative proteomics. Herein we describe the development of a novel maleyl-labeling-based approach for protein quantification. Under optimized conditions, maleic anhydride can serve as a highly efficient reagent to label the amino groups of tryptic peptides. Furthermore, “click chemistry” was successfully applied to obtain the second modification of maleylated peptides via thiol-Michael addition reaction. Accurate quantification was further achieved via the first or/and second step stable-isotope labeling in this study. Our data thus demonstrate that the maleyl-labeling-based method is simple, accurate, and reliable for quantitative proteomics. The developed method not only enables an enhanced sequence coverage of proteins by improving the identification of small and hydrophilic peptides, but also enables a controllable, successive, second derivatization of labeled peptides or proteins, and therefore holds a very promising potential for in-depth analysis of protein structures and dynamics.