Toshihiro Yoshimura, K. Kurogi, Ming-Cheh Liu, M. Suiko, Y. Sakakibara
{"title":"利用荧光标记技术分析s -亚硝基化蛋白质的蛋白质组学方法","authors":"Toshihiro Yoshimura, K. Kurogi, Ming-Cheh Liu, M. Suiko, Y. Sakakibara","doi":"10.2198/JELECTROPH.60.5","DOIUrl":null,"url":null,"abstract":"S-nitrosylation, a post-translational modification of the thiol group of cysteine residues by nitric oxide (NO), has emerged as a new mode of signal transduction and regulation of protein function. It has recently been shown that S-nitrosylation may result in various protein dysfunctions. However, an improved S-nitrosylation analysis method is needed to achieve high sensitivity and quantitative accuracy. We hypothesized that an analysis method using fluorescence dye could detect S-nitrosylated proteins at a higher sensitivity than that of the conventional method. In this study, we developed a procedure for analyzing S-nitrosylated proteins using CyDye (the CyDye switch method). This CyDye switch method for detecting S-nitrosylated proteins was developed based on the biotin-switch method for analyzing S-nitrosylated proteins. We analyzed NO donor-induced S-nitrosylated proteins in a model protein and at the cellular level. We demonstrated that this CyDye switch method could detect specific S-nitrosylated proteins using SDS-PAGE and mass spectrometry. Our results indicate that the optimized CyDye switch method is suitable for the detection of the post-translational S-nitrosylation of proteins.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"36 1","pages":"5-14"},"PeriodicalIF":0.0000,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"A proteomic approach for the analysis of S-nitrosylated proteins using a fluorescence labeling technique\",\"authors\":\"Toshihiro Yoshimura, K. Kurogi, Ming-Cheh Liu, M. Suiko, Y. Sakakibara\",\"doi\":\"10.2198/JELECTROPH.60.5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"S-nitrosylation, a post-translational modification of the thiol group of cysteine residues by nitric oxide (NO), has emerged as a new mode of signal transduction and regulation of protein function. It has recently been shown that S-nitrosylation may result in various protein dysfunctions. However, an improved S-nitrosylation analysis method is needed to achieve high sensitivity and quantitative accuracy. We hypothesized that an analysis method using fluorescence dye could detect S-nitrosylated proteins at a higher sensitivity than that of the conventional method. In this study, we developed a procedure for analyzing S-nitrosylated proteins using CyDye (the CyDye switch method). This CyDye switch method for detecting S-nitrosylated proteins was developed based on the biotin-switch method for analyzing S-nitrosylated proteins. We analyzed NO donor-induced S-nitrosylated proteins in a model protein and at the cellular level. We demonstrated that this CyDye switch method could detect specific S-nitrosylated proteins using SDS-PAGE and mass spectrometry. Our results indicate that the optimized CyDye switch method is suitable for the detection of the post-translational S-nitrosylation of proteins.\",\"PeriodicalId\":15059,\"journal\":{\"name\":\"Journal of capillary electrophoresis\",\"volume\":\"36 1\",\"pages\":\"5-14\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of capillary electrophoresis\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.2198/JELECTROPH.60.5\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of capillary electrophoresis","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.2198/JELECTROPH.60.5","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
A proteomic approach for the analysis of S-nitrosylated proteins using a fluorescence labeling technique
S-nitrosylation, a post-translational modification of the thiol group of cysteine residues by nitric oxide (NO), has emerged as a new mode of signal transduction and regulation of protein function. It has recently been shown that S-nitrosylation may result in various protein dysfunctions. However, an improved S-nitrosylation analysis method is needed to achieve high sensitivity and quantitative accuracy. We hypothesized that an analysis method using fluorescence dye could detect S-nitrosylated proteins at a higher sensitivity than that of the conventional method. In this study, we developed a procedure for analyzing S-nitrosylated proteins using CyDye (the CyDye switch method). This CyDye switch method for detecting S-nitrosylated proteins was developed based on the biotin-switch method for analyzing S-nitrosylated proteins. We analyzed NO donor-induced S-nitrosylated proteins in a model protein and at the cellular level. We demonstrated that this CyDye switch method could detect specific S-nitrosylated proteins using SDS-PAGE and mass spectrometry. Our results indicate that the optimized CyDye switch method is suitable for the detection of the post-translational S-nitrosylation of proteins.