High-Resolution Parallel Reaction Monitoring with Electron Transfer Dissociation for Middle-Down Proteomics: An Application to Study the Quantitative Changes Induced by Histone Modifying Enzyme Inhibitors and Activators

组蛋白 化学 乙酰化 组蛋白脱乙酰基酶 选择性反应监测 蛋白质组学 电子转移离解 定量蛋白质组学 生物化学 计算生物学 串联质谱法 组合化学 质谱法 色谱法 生物 DNA 基因
作者
Michael J. Sweredoski,Annie Moradian,Sonja Hess
出处
期刊:Methods in molecular biology 卷期号:: 61-69 被引量:5
标识
DOI:10.1007/978-1-4939-7201-2_4
摘要

With the advent of new methodologies, proteomics-based assays are increasingly used to study the efficacy of drugs on a molecular level. For these studies to be meaningful, the proteomics assays need to be sensitive, selective, accurate, and reproducible. This is often accomplished through a targeted approach, either using single or multiple reaction monitoring (SRM/MRM) or, more recently, parallel reaction monitoring (PRM). In PRM, the parallel detection of all product ions in a high-resolution mass spectrometer affords higher selectivity than SRM/MRM. PRM is thus better suited to analyze peptides larger than 2 kDa. Similar to SRM/MRM, PRM provides sensitive, accurate, and reproducible quantitative data. Here, we present a specific PRM method to characterize the effects of histone modifying enzyme drugs such as histone deacetylase inhibitors (HDAC) on the posttranslational modifications of histones, in a quantitative manner. More specifically, we characterize the heavily modified N-terminal tail of histone H3 after treatment with the HDAC inhibitor butyric acid, and monitor the acetylation and methylation events after treatment. To take most advantage of the multiply charged N-terminal histone peptides that are generated by an endoproteinase GluC-digestion, we use electron transfer dissociation (ETD) as the method of MS/MS fragmentation. This provides high sequence coverage for the modified peptides. The methodology is not limited to HDAC inhibitors, and can be used for any modifying enzyme. In fact, it can even be expanded beyond histone analyses. To give guidance for the development of a PRM assay, we present here HDAC inhibited H3 histone N-terminal tails as an example.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
oneyyy完成签到 ,获得积分10
刚刚
zjuszk完成签到 ,获得积分10
刚刚
刚刚
猫猫完成签到,获得积分10
1秒前
所所应助秋秋采纳,获得10
1秒前
桐桐应助Z777采纳,获得10
2秒前
Zheng关注了科研通微信公众号
2秒前
默默平文完成签到,获得积分10
2秒前
YRY完成签到 ,获得积分10
3秒前
3秒前
大清完成签到,获得积分10
3秒前
4秒前
雪白灵雁完成签到 ,获得积分10
4秒前
大气海露完成签到,获得积分10
4秒前
4秒前
精神的精神病完成签到,获得积分10
5秒前
6秒前
小马哥西北孤狼完成签到,获得积分10
6秒前
杪夏二八完成签到 ,获得积分10
6秒前
6秒前
Graham发布了新的文献求助10
7秒前
鹿芩完成签到,获得积分10
8秒前
iiii发布了新的文献求助10
8秒前
慧妞完成签到 ,获得积分10
8秒前
DL_LBK发布了新的文献求助10
8秒前
sugar完成签到,获得积分10
9秒前
思源应助科研顺利采纳,获得10
9秒前
罗亚亚发布了新的文献求助10
10秒前
小周小周完成签到,获得积分10
10秒前
TOM完成签到,获得积分10
10秒前
刘桔发布了新的文献求助10
10秒前
luo完成签到 ,获得积分10
11秒前
11秒前
酷酷雍完成签到 ,获得积分10
11秒前
LonelyCMA完成签到 ,获得积分10
11秒前
休思完成签到 ,获得积分10
11秒前
积极冷霜完成签到,获得积分10
12秒前
12秒前
怎么说来着完成签到,获得积分10
12秒前
12秒前
高分求助中
Sustainability in Tides Chemistry 1500
TM 5-855-1(Fundamentals of protective design for conventional weapons) 1000
Gerard de Lairesse : an artist between stage and studio 500
Digging and Dealing in Eighteenth-Century Rome 500
Queer Politics in Times of New Authoritarianisms: Popular Culture in South Asia 500
Livre et militantisme : La Cité éditeur 1958-1967 500
Retention of title in secured transactions law from a creditor's perspective: A comparative analysis of selected (non-)functional approaches 500
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3063367
求助须知:如何正确求助?哪些是违规求助? 2718227
关于积分的说明 7457962
捐赠科研通 2364609
什么是DOI,文献DOI怎么找? 1253459
科研通“疑难数据库(出版商)”最低求助积分说明 608647
版权声明 596606