Protein turnover measurement using selected reaction monitoring-mass spectrometry (SRM-MS).
Protein turnover measurement using selected reaction monitoring-mass spectrometry (SRM-MS).
复制标题
DOI：
10.1098/rsta.2015.0362
复制
发表时间：
2016-10-28
期刊：
影响因子：
--
通讯作者：
Beynon RJ
中科院分区：
文献类型：
--
作者：
Holman SW;Hammond DE;Simpson DM;Waters J;Hurst JL;Beynon RJ
蛋白质周转代表了细胞功能的重要机制，与许多疾病状态相关的蛋白质合成和降解不受管制。因此，用蛋白质组学策略来测量高可信度的周转率对于理解许多生物过程至关重要。在本研究中，更广泛使用的非靶向前体离子信号强度（MS1）定量方法与选择性反应监测（SRM）（一种记录特定肽数据的数据采集策略）进行了比较，以确定使用靶向定量方法是否可以获得更好的定量数据。以小鼠肝脏为模型系统，采用MS1和SRM两种定量策略对四种三羧酸循环蛋白进行周转测定。SRM在测量的灵敏度和选择性方面优于MS1，可以更有信心地确定蛋白质周转率。SRM数据是使用更便宜和更广泛使用的串联四极杆质谱仪获得的，这使得该方法比MS1量化方法更容易被更多的研究人员使用，MS1量化方法在高质量分辨率仪器上表现最好。SRM采集非常适合于测量数十种蛋白质周转的集中研究，使其适用于确定蛋白质复合物的动力学和完整的代谢途径。本文是“定量质谱”专题的一部分。
Protein turnover represents an important mechanism in the functioning of cells, with deregulated synthesis and degradation of proteins implicated in many diseased states. Therefore, proteomics strategies to measure turnover rates with high confidence are of vital importance to understanding many biological processes. In this study, the more widely used approach of non-targeted precursor ion signal intensity (MS1) quantification is compared with selected reaction monitoring (SRM), a data acquisition strategy that records data for specific peptides, to determine if improved quantitative data would be obtained using a targeted quantification approach. Using mouse liver as a model system, turnover measurement of four tricarboxylic acid cycle proteins was performed using both MS1 and SRM quantification strategies. SRM outperformed MS1 in terms of sensitivity and selectivity of measurement, allowing more confident determination of protein turnover rates. SRM data are acquired using cheaper and more widely available tandem quadrupole mass spectrometers, making the approach accessible to a larger number of researchers than MS1 quantification, which is best performed on high mass resolution instruments. SRM acquisition is ideally suited to focused studies where the turnover of tens of proteins is measured, making it applicable in determining the dynamics of proteins complexes and complete metabolic pathways. This article is part of the themed issue ‘Quantitative mass spectrometry’.
登录
查看更多内容
DOI：
10.1074/mcp.m113.034769
发表时间：
2014-01
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Hebert AS;Richards AL;Bailey DJ;Ulbrich A;Coughlin EE;Westphall MS;Coon JJ
通讯作者：
Coon JJ
影响因子：
3.4
作者：
Brownridge, Philip;Lawless, Craig;Payapilly, Aishwarya B.;Lanthaler, Karin;Holman, Stephen W.;Harman, Victoria M.;Grant, Christopher M.;Beynon, Robert J.;Hubbard, Simon J.
通讯作者：
Hubbard, Simon J.
影响因子：
8.8
作者：
Christiano R;Nagaraj N;Fröhlich F;Walther TC
通讯作者：
Walther TC
DOI：
10.1016/j.bbagen.2005.12.023
发表时间：
2006-05-01
影响因子：
3
作者：
Busch, Robert;Kim, Yoo-Kyeong;Hellerstein, Marc K.
通讯作者：
Hellerstein, Marc K.
DOI：
10.1074/mcp.m114.047407
发表时间：
2015-07
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Beck S;Michalski A;Raether O;Lubeck M;Kaspar S;Goedecke N;Baessmann C;Hornburg D;Meier F;Paron I;Kulak NA;Cox J;Mann M
通讯作者：
Mann M