Comparative evaluation of extraction methods for apoplastic proteins from maize leavesReport as inadecuate




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Plant Methods

, 7:48

First Online: 22 December 2011Received: 01 November 2011Accepted: 22 December 2011

Abstract

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.

KeywordsApoplast liquid chromatography mass spectrometry maize proteome analysis two-dimensional gel electrophoresis AbbreviationsLC-MS-MSliquid chromatography tandem mass spectrometry

1-DEone-dimensional gel electrophoresis

2-DEtwo-dimensional gel electrophoresis

Electronic supplementary materialThe online version of this article doi:10.1186-1746-4811-7-48 contains supplementary material, which is available to authorized users.

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Author: Katja Witzel - Muhammad Shahzad - Andrea Matros - Hans-Peter Mock - Karl H Mühling

Source: https://link.springer.com/







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