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A strategy for the enrichment and characterization of disulfide bond-contained proteins from Chinese cobra (Naja atra) venom |
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| Authors: | Yuanqing Wei Ting Liu Binru Zheng Yilin Song Shengsong Wang Mojuan Zheng Yanling Xu Yumei Chi Ming Zhao Jin-ao Duan Shuying Han Rui Liu |
| Institution: | 1. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023 P. R. China
Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing, 210023 P. R. China Jiangsu Key Laboratory of Research and Development in Marine Bio-resource Pharmaceutics, Nanjing University of Chinese Medicine, Nanjing, 210023 P. R. China These authors contributed equally to this work.;2. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023 P. R. China These authors contributed equally to this work.;3. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023 P. R. China;4. Anhui Qimen Institute of Snakebite, Huangshan, 245000 P. R. China;5. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023 P. R. China Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing, 210023 P. R. China |
| Abstract: | A new strategy combined gold-coated magnetic nanocomposites assisted enrichment with mass spectrometry was developed for the characterization of disulfide bond-contained proteins from Chinese cobra (Naja atra) venom. In this work, core-shell nanocomposites were synthesized by the seed-mediated growth method and used for the enrichment of snake venom proteins containing disulfide bonds. A total of 3545 tryptic digested peptides derived from 96 venom proteins in Naja atra venom were identified. The venom proteins comprised 14 toxin families including three-finger toxins, phospholipase A2, snake venom metalloproteinase, cobra venom factor, and so forth. Extra 16 venom proteins were detected exclusively in the nanocomposites set, among which 11 venom proteins were from the three-finger toxins family. In the present study, the proposed simple and efficient protocol replaced the tedious and laborious technologies commonly used for pre-separating crude snake venom, suggesting widely implementation in low-abundance or trace disulfide bond-contained proteins or peptides characterization. |
| Keywords: | enrichment mass spectrometry Naja atra nanoparticles venom proteome |