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

Yılmaz C., İşcan M.

Current Proteomics, cilt.18, sa.5, ss.742-752, 2021 (SCI-Expanded)

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 18 Sayı: 5
  • Basım Tarihi: 2021
  • Doi Numarası: 10.2174/1570164618999201209221340
  • Dergi Adı: Current Proteomics
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Biotechnology Research Abstracts, Chemical Abstracts Core, EMBASE
  • Sayfa Sayıları: ss.742-752
  • Van Yüzüncü Yıl Üniversitesi Adresli: Evet

Özet

Aim: 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.