Chloroplast isolation and affinity chromatography for enrichment of low-abundant proteins in complex proteomes
- Author(s)
- Roman G Bayer, Simon Stael, Markus Teige
- Abstract
Detailed knowledge of the proteome is crucial to advance the biological sciences. Low-abundant proteins are of particular interest to many biologists as they include, for example those proteins involved in signal transduction. Recent technological advances resulted in a tremendous increase in protein identification sensitivity by mass spectrometry (MS). However, the dynamic range in protein abundance still forms a fundamental problem that limits the detection of low-abundant proteins in complex proteomes. These proteins will typically escape detection in shotgun MS experiments due to the presence of other proteins at an abundance several-fold higher in order of magnitude. Therefore, specific enrichment strategies are required to overcome this technical limitation of MS-based protein discovery. We have searched for novel signal transduction proteins, more specifically kinases and calcium-binding proteins, and here we describe different approaches for enrichment of these low-abundant proteins from isolated chloroplasts from pea and Arabidopsis for subsequent proteomic analysis by MS. These approaches could be extended to include other signal transduction proteins and target different organelles.
- Organisation(s)
- Department of Biochemistry and Cell Biology
- External organisation(s)
- Ghent University , Universität für Bodenkultur Wien
- Volume
- 1295
- Pages
- 211-223
- No. of pages
- 13
- DOI
- https://doi.org/10.1007/978-1-4939-2550-6_16
- Publication date
- 2015
- Peer reviewed
- Yes
- Austrian Fields of Science 2012
- 106031 Plant physiology
- Keywords
- ASJC Scopus subject areas
- Genetics, Molecular Biology
- Portal url
- https://ucris.univie.ac.at/portal/en/publications/chloroplast-isolation-and-affinity-chromatography-for-enrichment-of-lowabundant-proteins-in-complex-proteomes(482d597e-6265-4209-8a31-23c5d4878ee5).html