Identification of Plant Nuclear Proteins Based on a Combination of Flow Sorting, SDS-PAGE, and LC-MS/MS Analysis
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- Cell cycle, Flow cytometry, Gel electrophoresis, In-gel digestion, Mass spectrometry, Nuclear proteome, Plant nucleus, Protein analysis,
- MeSH
- Electrophoresis, Polyacrylamide Gel MeSH
- Nuclear Proteins analysis MeSH
- Hordeum cytology metabolism MeSH
- Gas Chromatography-Mass Spectrometry MeSH
- Proteomics methods MeSH
- Flow Cytometry MeSH
- Plant Proteins analysis MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Nuclear Proteins MeSH
- Plant Proteins MeSH
In the plant nucleus, the majority of cellular DNA content is stored and maintained. This makes this highly specialized organelle the major coordinator of almost all essential processes in plant cells such as transcription, DNA replication, and repair. None of these biological pathways can be fully understood without a comprehensive characterization of nuclear proteins. Nevertheless, the interest of the proteomic community in the plant nuclear proteome has been very limited so far. This is probably due to the high integrity of plant cell, presence of many interfering metabolites, and considerable endogenous proteolytic activity which make the sample preparation problematic. Hereby, we describe a novel protocol for the high-throughput plant nuclear protein identification that combines a flow cytometric sorting of formaldehyde-fixed nuclei with protein and peptide separation and their subsequent LC-MS/MS analysis.
References provided by Crossref.org
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