期刊
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS
卷 1854, 期 5, 页码 460-468出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbapap.2015.01.006
关键词
ERLIC; Proteomics; Phosphoproteomics; Multidimensional separation; Clinical research
资金
- Ministry for Innovation, Science and Research of the Federal State of North Rhine-Westphalia
- Free and Hanseatic City of Hamburg
- Federal Ministry of Health
Quantitative proteomics and phosphoproteomics have become key disciplines in understanding cellular processes. Fundamental research can be done using cell culture providing researchers with virtually infinite sample amounts. In contrast, clinical, pre-clinical and biomedical research is often restricted to minute sample amounts and requires an efficient analysis with only micrograms of protein. To address this issue, we generated a highly sensitive workflow for combined LC-MS-based quantitative proteomics and phosphoproteomics by refining an ERLIC-based 2D phosphoproteomics workflow into an ERLIC-based 3D workflow covering the global proteome as well. The resulting 3D strategy was successfully used for an in-depth quantitative analysis of both, the proteome and the phosphoproteome of murine cytomegalovirus-infected mouse fibroblasts, a model system for host cell manipulation by a virus. In a 2-plex SILAC experiment with 150 mu g of a tryptic digest per condition, the 3D strategy enabled the quantification of similar to 75% more proteins and even similar to 134% more peptides compared to the 2D strategy. Additionally, we could quantify similar to 50% more phosphoproteins by non-phosphorylated peptides, concurrently yielding insights into changes on the levels of protein expression and phosphorylation. Beside its sensitivity, our novel three-dimensional ERLIC-strategy has the potential for semi-automated sample processing rendering it a suitable future perspective for clinical, pre-clinical and biomedical research. (C) 2015 Published by Elsevier B.V.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据