Projects per year
Systems biological approaches to study the Arabidopsis thaliana circadian clock have mainly focused on transcriptomics while little is known about the proteome, and even less about posttranslational modifications. Evidence has emerged that posttranslational protein modifications, in particular phosphorylation, play an important role for the clock and its output. Phosphoproteomics is the method of choice for a large-scale approach to gain more knowledge about rhythmic protein phosphorylation. Recent plant phosphoproteomics publications have identified several thousand phosphopeptides. However, the methods used in these studies are very labor-intensive and therefore not suitable to apply to a well-replicated circadian time series. To address this issue, we present and compare different strategies for sample preparation for phosphoproteomics that are compatible with large numbers of samples. Methods are compared regarding number of identifications, variability of quantitation, and functional categorization. We focus on the type of detergent used for protein extraction as well as methods for its removal. We also test a simple two-fraction separation of the protein extract.
|Title of host publication||Methods in Enzymology|
|Subtitle of host publication||Membrane Proteins—Engineering, Purification and Crystallization|
|Number of pages||27|
|Publication status||Published - 2015|
- Circadian clock
- Mass spectrometry
- Protein extraction
FingerprintDive into the research topics of 'Sample preparation for phosphoproteomic analysis of circadian time series in Arabidopsis thaliana'. Together they form a unique fingerprint.
- 3 Finished
Does an Ancient Circadian Clock control transcriptional rhythms using a non transcriptional oscillator
Millar, A. & Le Bihan, T.
1/10/12 → 31/01/16
R42038 Wellcome Trust Four Year PhD Studentship Ms Johanna Krahmer
1/10/11 → 30/09/15
SynthSys; formerly CSBE: Centre for Systems Biology at Edinburgh
Millar, A., Beggs, J., Ghazal, P., Goryanin, I., Hillston, J., Plotkin, G., Tollervey, D., Walton, A. & Robertson, K.
8/01/07 → 31/12/12
Proteomic data from "Sample preparation for phosphoproteomic analysis of circadian time series in Arabidopsis thaliana"
Krahmer, J. (Creator), Martin, S. (Creator), Hindle, M. (Creator), Le Bihan, T. (Creator) & Millar, A. (Creator), PRIDE database hosted by European Bioinformatics Institute, EBI, 2016
DOI: 10.7488/ba4de74b-7d98-48c8-b587-c43f44dc37c9, http://www.ebi.ac.uk/pride/archive/projects/PXD001134