17th International Mass Spectrometry Conference :: Prague, 2006
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|Session:||LATE-BREAKING/Proteomics: New Methods|
|Presentation date:||Tue, Aug 29, 2006|
|Presentation time:||09:50 – 11:20|
Andrea Matros1, Steffen Amme1, Hans-Peter Mock1, Jim Langridge21 IPK-Gatersleben, Gatersleben, Germany
Correspondence address: Andrea Matros, IPK-Gatersleben, Applied Biochemistry, Corrensstr., 3, Gatersleben, 06466 Germany.
Keywords: Chromatography, Liquid -, Nanoscale Capillary; Electrophoresis, 2D; Protein Identification; Sample Preparation.
Novel aspect: proteome study on cold stress response, LC-based separation combined with ESI-Q-TOF MS
Environmental influences such as high light, drought, high or low temperature and salinity affect growth and yield of crop plants by leading to altered gene and protein expression, metabolic changes, and growth retardation. How such environmental stimuli are perceived and trigger the complex defensive and adaptive signalling networks, and how these events result in resistance/tolerance is of major practical interest. A proteome study based on 2-D gel electrophoresis was performed to analyse the cold stress response of Arabidopsis plants with an emphasis to monitor the overall changes in the protein complement. Two different temperature regimes were used (6°C and 10°C) and plants were exposed to cold stress for one week. To monitor gradual changes in the response to cold stress exposure we carried out a comparison of protein patterns by using DIGE technology. A number of spots displaying at least two-fold changes in relative abundance were chosen for identification. Spots were excised from 2-D gels, tryptically digested and analysed by MALDI-TOF MS or ESI-Q-TOF MS. Proteins displaying significant changes in abundance included RNA-binding protein CP29, a glycin-rich protein, carbohydrate metabolism-associated proteins, dehydrins and low-temperature-induced protein 78. Many of the proteins identified have previously been described in the context of cold stress responses, indicating the validity of our proteome approach for further in depth studies. Taking advantage of recent developments in proteome analysis samples from these studies were also analysed by a label free quantification method using LC-based separation combined with ESI-Q-TOF MS. This novel approach confirmed earlier results obtained by using DIGE technology but also lead to the identification of a number of additional proteins with changed expression pattern after cold stress exposure.