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An optimized method to extract poplar leaf proteins for two‐dimensional gel electrophoresis guided by analysis of polysaccharides and phenolic compounds
Author(s) -
Szuba Agnieszka,
Wojakowska Anna,
Lorenc–Plucińska Gabriela
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201300223
Subject(s) - polyphenol , polysaccharide , cultivar , extraction (chemistry) , starch , chemistry , acetone , phenol extraction , chromatography , phenol , food science , botany , biology , biochemistry , gene , organic chemistry , antioxidant , rna
Commonly used methods for protein extraction from plant leaves, such as extraction with phenol or a combination of trichloroacetic acid and acetone, were ineffective for four tested cultivars of poplar. Moreover, multiple protocols for 2 DE of the extracted proteins gave different results when protein profiles of relatively closely related plants were compared. Given that polycyclic compounds strongly hinder 2 DE , we analyzed the impact of polyphenols and polysaccharides present in the plant tissues used for protein extraction, on the quality of 2 DE protein profiles. Analysis of content of polyphenols and polysaccharides in leaves of poplar cultivars showed that even small differences in concentrations of analyzed metabolites accompany large differences between poplar cultivars when considering the susceptibility of samples to protein extraction for 2 DE . High‐quality 2 DE results were correlated with decreased amounts of polyphenols. Additional analysis using MS / MS suggested that only levels of total phenolics affected the results of 2 DE . Soluble total nonstructural carbohydrates also had a negative effect, but the level of starch was not important. Finally, we present an optimized method for extraction of proteins from poplar leaves, which enables reliable comparative analysis of four different poplar cultivars, that is, “Eridano,” “Villafranca,” “NE‐42,” and “Luisa Avanzo,” which have not yet been used for the proteomic studies.