Integrative Chemical Proteomics and Cell Biology Methods to Study Endocytosis and Vesicular Trafficking in Arabidopsis
We present a comprehensive approach combining proteomics and cell biology to study vesicular trafficking in plants. Within this approach, we exploit chemical compounds inhibiting particular vesicular trafficking events in plant cells. Treatment of plants with these relatively specific inhibitors results in intracellular accumulation of proteins being transported by vesicles as well as in a change in abundance of regulatory proteins. Such pharmacological inhibition allows for identification of key proteins, and for further detailed functional investigation using cell biological, molecular biological, and biochemical methods used for validation of proteomic results.
Key wordsEndocytosis Vesicular trafficking Proteomics Plants 2-D LC/MSMS 2-D electrophoresis Inhibitors Brefeldin A Wortmannin LY294002
This work was supported by the grant LO1204 from the National Program of Sustainability I to the Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, by Czech Republic Operational Program Education for Competitiveness—European Social Fund (project CZ.1.07/2.3.00/20.0165), by Genomics for Southern Crop Stress and Disease, USDA CSREES 2009-34609-20222, and by grant No. NIH INBRE P20GM103476.
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