Abstract

Aims: This study aimed to generate an improved method of protein extraction and purification from plant tissues containing very high amounts of phenolic compounds and other interfering 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 enhanced abilities to produce higher purity of samples compatible for high precision molecular systems and analysis.

Methods: The new method was compared with the other two for their efficiencies in classical SDS- 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 concentrations 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.

Keywords: 2-DE, IEF, pinus, protein chip, proteomics, SDS, protein extraction.

Graphical Abstract