ESNOQ, Proteomic Quantification of Endogenous S-Nitrosation

	ESNOQ, Proteomic Quantification of Endogenous S-Nitrosation
	Zhou, XX; Han, PW; Li, JM; Zhang, X; Huang, B; Ruan, HQ; Chen, C
刊名	PLOS ONE
	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
内容类型	期刊论文
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.