FluoroTRAQ: Quantitative Analysis of Protein S-Nitrosylation through Fluorous Solid-Phase Extraction Combining with iTRAQ by Mass Spectrometry
| Autor: | Haojie Lu, Yaoyao Xu, Guoli Wang, Huimin Bao, Ying Zhang, Cheng Zhang, Caiyun Fang |
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| Rok vydání: | 2020 |
| Předmět: |
Proteomics
chemistry.chemical_classification Protein nitrosylation Solid Phase Extraction Proteins Nitric Oxide Mass spectrometry Mass Spectrometry Analytical Chemistry Amino acid Kinetics chemistry Biochemistry Human Umbilical Vein Endothelial Cells Humans Solid phase extraction Signal transduction Protein Processing Post-Translational Quantitative analysis (chemistry) Cysteine |
| Zdroj: | Analytical Chemistry. 92:15317-15322 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.0c01706 |
| Popis: | S-Nitrosylation is an important post-translational modification that occurs on cysteine amino acid and regulates signal transduction in diverse cell processes. Dysregulation of protein nitrosylation has shown close association with cardiovascular and neurological diseases, thus demanding further precise and in-depth understanding. Mass spectrometry-based proteomics has been the method of choice for analyzing S-nitrosylated (SNO-) proteins. However, due to their extremely low expression level and rapid turnover rate, quantitative analysis of the S-nitrosylation at the proteomic level remains challenging. Herein, we developed a novel approach termed FluoroTRAQ, which combined the fluorous solid-phase extraction of SNO-peptides and iTRAQ labeling for the quantitative analysis of the SNO-proteome with high sensitivity and specificity. This new analytical strategy was subsequently applied to examine the dynamic SNO-proteome changes of human umbilical vein endothelial cells upon in vitro S-nitrosoglutathione induction. Our data identified a number of novel SNO-proteins and revealed their temporal modulation as validated by biotin switch assay. Our study offered a practical approach for quantitative analysis of protein S-nitrosylation. |
| Databáze: | OpenAIRE |
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