Department of Genome Sciences, University of Washington, Seattle, Washington 98109-4717, United States.
Anal Chem. 2011 Nov 15;83(22):8403-10. doi: 10.1021/ac2017053. Epub 2011 Oct 27.
Stable isotope labeling by amino acids in cell culture (SILAC) is a versatile tool in proteomics that has been used to explore protein turnover on a large scale. However, these studies pose a significant undertaking that can be greatly simplified through the use of computational tools that automate the data analysis. While SILAC technology has enjoyed rapid adoption through the availability of several software tools, algorithms do not exist for the automated analysis of protein turnover data generated using SILAC technology. Presented here is a software tool, SILACtor, designed to trace and compare SILAC-labeled peptides across multiple time points. SILACtor is used to profile protein turnover rates for more than 500 HeLa cell proteins using a SILAC label-chase approach. Additionally, SILACtor contains a method for the automated generation of accurate mass and retention time inclusion lists that target peptides of interest showing fast or slow turnover rates relative to the other peptides observed in the samples. SILACtor enables improved protein turnover studies using SILAC technology and also provides a framework for features extensible to comparative SILAC analyses and targeted methods.
稳定同位素标记的氨基酸在细胞培养(SILAC)是蛋白质组学中的一种通用工具,已被用于大规模探索蛋白质周转。然而,这些研究是一项重大的任务,可以通过使用自动化数据分析的计算工具大大简化。尽管 SILAC 技术通过几种软件工具的可用性得到了快速采用,但目前还没有用于自动分析使用 SILAC 技术生成的蛋白质周转数据的算法。本文介绍了一种软件工具 SILACtor,用于跟踪和比较多个时间点的 SILAC 标记肽。SILACtor 用于通过 SILAC 标记追踪方法对超过 500 个 HeLa 细胞蛋白的蛋白质周转率进行分析。此外,SILACtor 还包含一种自动生成准确质量和保留时间包含列表的方法,该方法针对相对于样品中观察到的其他肽具有快速或慢速周转率的感兴趣肽。SILACtor 可使用 SILAC 技术改进蛋白质周转研究,并且还为可扩展到比较 SILAC 分析和靶向方法的功能提供了框架。