Brownridge Philip J, Harman Victoria M, Simpson Deborah M, Beynon Robert J
Protein Function Group, Institute of Integrative Biology, University of Liverpool, Liverpool, UK.
Methods Mol Biol. 2012;893:267-93. doi: 10.1007/978-1-61779-885-6_18.
In addition to protein identification, protein quantification is becoming a key output of proteomic experiments. Although relative quantification techniques are more commonplace and central to discovery proteomics, most assays require absolute quantification. The growth in systems biology has also increased the demand for absolute protein abundance values for input into models. QconCATs are created by concatenating peptide sequences taken from the target proteins into artificial proteins. The QconCAT acts as a source of internal standards and enables parallel absolute quantification of multiple proteins. QconCATs are typically applied in targeted proteomic workflows and so benefit from the greater sensitivity and wider dynamic range of these approaches. In this chapter, we discuss the design, construction, expression, and deployment of a QconCAT and the resulting experiments required for multiplex absolute quantification.
除了蛋白质鉴定外，蛋白质定量正成为蛋白质组学实验的关键产出。尽管相对定量技术在发现蛋白质组学中更为常见且是核心内容，但大多数检测需要绝对定量。系统生物学的发展也增加了对用于模型输入的绝对蛋白质丰度值的需求。QconCAT是通过将取自目标蛋白质的肽序列连接成人工蛋白质而创建的。QconCAT作为内标来源，能够对多种蛋白质进行平行绝对定量。QconCAT通常应用于靶向蛋白质组学工作流程，因此受益于这些方法更高的灵敏度和更宽的动态范围。在本章中，我们将讨论QconCAT的设计、构建、表达和应用以及多重绝对定量所需的后续实验。
