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Extraction and Proteome Analysis of Starch Granule-Associated Proteins in Mature Wheat Kernel (Triticum aestivum L.)

Starch consists of the two glucose polymers, amylose and amylopectin, and is deposited as semicrystalline granules inside plastids. The starch granule proteome is particularly challenging to study due to the amount of interfering compounds (sugars, storage proteins), the very low starch granule-associated protein content and also the dynamic range of abundant proteins. Here we present the protocol for extraction and 2-DE of wheat starch granule-associated proteins whose most important steps are: (i) washing and sonication to remove interfering compounds (storage proteins) from the surface of the granules, (ii) scanning electron microscopy (SEM) observations to monitor purification and granules swelling, (iii) appropriate protein extraction and solubilization to obtain enough proteins for Coomassie blue staining and proteomic analysis. Our objective was to minimize the amount of contamination by storage proteins and to preserve the structure of the starch and of starch-associated proteins and to maximize t...

Authors:   Emmanuelle Bancel; Hélène Rogniaux; Clément Debiton; Christophe Chambon; Gérard Branlard; Emmanuelle Bancel; Hélène Rogniaux
Journal:   Journal of Proteome Research
Year:   2010
DOI:   10.1021/pr9010525
Publication date:   01-06-2010

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