Comparative evaluation of extraction methods for apoplastic proteins from maize leaves

Plant Methods - Tập 7 - Trang 1-11 - 2011
Katja Witzel1,2, Muhammad Shahzad1, Andrea Matros3, Hans-Peter Mock3, Karl H Mühling1
1Institute of Plant Nutrition and Soil Science, Christian Albrechts University, Kiel, Germany
2Leibniz Institute of Vegetable and Ornamental Crops, Großbeeren, Germany
3Leibniz Institute of Plant Genetics and Crop Plant Research, Gatersleben, Germany

Tóm tắt

Proteins in the plant apoplast are essential for many physiological processes. We have analysed and compared six different infiltration solutions for proteins contained in the apoplast to recognize the most suitable method for leaves and to establish proteome maps for each extraction. The efficiency of protocols was evaluated by comparing the protein patterns resolved by 1-DE and 2-DE, and revealed distinct characteristics for each infiltration solution. Nano-LC-ESI-Q-TOF MS analysis of all fractions was applied to cover all proteins differentially extracted by infiltration solutions and led to the identification of 328 proteins in total in apoplast preparations. The predicted subcellular protein localisation distinguished the examined infiltration solutions in those with high or low amounts of intracellular protein contaminations, and with high or low quantities of secreted proteins. All tested infiltration solution extracted different subsets of proteins, and those implications on apoplast-specific studies are discussed.

Tài liệu tham khảo

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