Journal
ANALYTICAL CHEMISTRY
Volume 89, Issue 16, Pages 8259-8265Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b01120
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Funding
- National Natural Science Foundation of China [21275077]
- National Basic Research Program of China [2016YFC0903000, 2013CB910903, 2012CB910601]
- Tianjin Municipal Science and Technology Commission [14JCYBJC24000]
- Talent Excellence Program from Tianjin Medical University
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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.
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