An optimized method to extract poplar leaf proteins for twodimensional gel electrophoresis guided by analysis of polysaccharides and phenolic compounds
PBN-AR
Instytucja
Instytut Dendrologii Polskiej Akademii Nauk
Informacje podstawowe
Główny język publikacji
EN
Czasopismo
ELECTROPHORESIS
ISSN
0173-0835
EISSN
Wydawca
URL
Rok publikacji
2013
Numer zeszytu
Strony od-do
3234-3243
Numer tomu
34
Identyfikator DOI
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Autorzy
Pozostali autorzy
+ 1
Słowa kluczowe
EN
2DE, Leaves, Polycyclic compounds, Poplar cultivars, Protein isolation
Streszczenia
Język
EN
Treść
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 2DE of the extracted proteins gave different results when protein profiles of relatively closely related plants were compared. Given that polycyclic compounds strongly hinder 2DE, we analyzed the impact of polyphenols and polysaccharides present in the plant tissues used for protein extraction, on the quality of 2DE 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 2DE. High-quality 2DE results were correlated with decreased amounts of polyphenols. Additional analysis using MS/MS suggested that only levels of total phenolics affected the results of 2DE. 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.
Inne
System-identifier
PX-56b461df8106eb71826e31d9
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