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


Yılmaz C., İŞCAN M.

CURRENT PROTEOMICS, vol.18, no.5, pp.742-752, 2021 (Journal Indexed in SCI) identifier identifier

  • Publication Type: Article / Article
  • Volume: 18 Issue: 5
  • Publication Date: 2021
  • Doi Number: 10.2174/1570164618999201209221340
  • Title of Journal : CURRENT PROTEOMICS
  • Page Numbers: pp.742-752
  • Keywords: 2-DE, IEF, pinus, protein chip, proteomics, SDS, PHENOLIC-COMPOUNDS, 2-DIMENSIONAL ELECTROPHORESIS, GEL-ELECTROPHORESIS, NEEDLES, PINE, CHALLENGES, RESPONSES

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.