A Combined Method of Protein Extraction From Unorthodox Plant Samples for Proteomics

Abstract

Aim: This study aimed to generate an improved method of protein extraction and purifi-cation from plant tissues containing very high amounts of phenolic compounds and other interfer-ing biomolecules. Background: Protein extraction at proteomic studies on some plant species, including conifers, is challenging, and the yield and quality are unpredictable. Objective: Two popular protocols were combined with each other to construct a novel one with en-hanced abilities to produce higher purity of samples compatible for high precision molecular sys-tems and analysis. Methods: The new method was compared with the other two for their efficiencies in classical SD-S-PAGE, 2-DE and capillary chromatography applications. Results: All three methods were comparable in SDS-PAGE procedure; however, only the new method created acceptable gel images in 2-DE. Bioanalyzer results, also, demonstrated that the new method provided protein samples pure enough to be used in capillary chromatography with 2 times more peaks in electropherograms with lower noise and higher total relative protein concentra-tions closest to the applied amount. Conclusion: The new combined method is a successful alternative for plant proteomicists with higher yield and quality of proteins from recalcitrant tissues. Other: The new method could be preferred, especially, for high-tech, sensitive proteomic analysis.