ESNOQ, Proteomic Quantification of Endogenous S-Nitrosation
文献类型：期刊论文
| 作者 | Zhou, XX; Han, PW; Li, JM; Zhang, X; Huang, B; Ruan, HQ; Chen, C |
| 刊名 | PLOS ONE
 |
| 出版日期 | 2010 |
| 卷号 | 5期号:3页码:e10015-e10015 |
| 通讯作者 | Zhou, XX (reprint author), Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing 100080, Peoples R China.,changchen@moon.ibp.ac.cn |
| 英文摘要 | S-nitrosation is a post-translational protein modification and is one of the most important mechanisms of NO signaling. Endogenous S-nitrosothiol (SNO) quantification is a challenge for detailed functional studies. Here we developed an ESNOQ (Endogenous SNO Quantification) method which combines the stable isotope labeling by amino acids in cell culture (SILAC) technique with the detergent-free biotin-switch assay and LC-MS/MS. After confirming the accuracy of quantification in this method, we obtained an endogenous S-nitrosation proteome for LPS/IFN-gamma induced RAW264.7 cells. 27 S-nitrosated protein targets were confirmed and using our method we were able to obtain quantitative information on the level of S-nitrosation on each modified Cys. With this quantitative information, over 15 more S-nitrosated targets were identified than in previous studies. Based on the quantification results, we found that the S-nitrosation levels of different cysteines varied within one protein, providing direct evidence for differences in the sensitivity of cysteine residues to reactive nitrosative stress and that S-nitrosation is a site-specific modification. Gene ontology clustering shows that S-nitrosation targets in the LPS/IFN-gamma induced RAW264.7 cell model were functionally enriched in protein translation and glycolysis, suggesting that S-nitrosation may function by regulating multiple pathways. The ESNOQ method described here thus provides a solution for quantification of multiple endogenous S-nitrosation events, and makes it possible to elucidate the network of relationships between endogenous S-nitrosation targets involved in different cellular processes. |
| 学科主题 | Science & Technology - Other Topics |
| 类目[WOS] | Multidisciplinary Sciences |
| 关键词[WOS] | NITROSYLATED PROTEINS ; MASS-SPECTROMETRY ; CELL-CULTURE ; NITRIC-OXIDE ; AMINO-ACIDS ; SILAC ; IDENTIFICATION ; SITES |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000276419900015 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/966]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Zhou, XX,Han, PW,Li, JM,et al. ESNOQ, Proteomic Quantification of Endogenous S-Nitrosation[J]. PLOS ONE,2010,5(3):e10015-e10015. |
| APA | Zhou, XX.,Han, PW.,Li, JM.,Zhang, X.,Huang, B.,...&Chen, C.(2010).ESNOQ, Proteomic Quantification of Endogenous S-Nitrosation.PLOS ONE,5(3),e10015-e10015. |
| MLA | Zhou, XX,et al."ESNOQ, Proteomic Quantification of Endogenous S-Nitrosation".PLOS ONE 5.3(2010):e10015-e10015. |
入库方式： OAI收割
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。