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Abstract

Comparative evaluation of extraction methods for apoplastic proteins from maize leaves.

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.



Witzel, K.; Shahzad, M.; Matros, A.; Mock, H-P.; Mühling KH. 2011. Comparative evaluation of extraction methods for apoplastic proteins from maize leaves. Plant Methods 7 (48), doi:10.1186/1746-4811-7-48