使用二氧化钛微柱和无标记定量的定量磷酸化蛋白质组学

Quantitative Phosphoproteomic Using Titanium Dioxide Micro-Columns and Label-Free Quantitation.

作者信息

Barrios-Llerena Martin E, Le Bihan Thierry

机构信息

Proteomics and Mass Spectrometry, Bioscience Core Labs, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia.

Synthetic and Systems Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, UK.

出版信息

Methods Mol Biol. 2019;1977:35-42. doi: 10.1007/978-1-4939-9232-4_4.

DOI:10.1007/978-1-4939-9232-4_4

PMID:30980321

Abstract

Phosphorylation events are important during cellular function. Analysis of phosphorylation in complex samples has been extensively studied using large-scale phosphopeptide enrichment methods. Quantitative analysis of the enriched phosphopeptides is subsequently performed using label-based methodologies (e.g., SILAC, iTRAQ, and others). Here we describe the protocol for the quantitative analysis of phosphopeptides, enriched with titanium dioxide micro-column, using an intensity-based label-free quantitation.

摘要

磷酸化事件在细胞功能过程中很重要。使用大规模磷酸肽富集方法对复杂样品中的磷酸化进行分析已得到广泛研究。随后使用基于标记的方法（例如，稳定同位素标记氨基酸法、同位素标记相对和绝对定量法等）对富集的磷酸肽进行定量分析。在此，我们描述了使用基于强度的无标记定量法对用二氧化钛微柱富集的磷酸肽进行定量分析的方案。