Investigation of plant latex proteins of genus Euphorbia L. by two-dimensional gel electrophoresis

Two dimensional gel electrophoresis (2-DE), described by O'Farrell, represents one of the most appropriate techniques for separation of complex protein pattern, and combines IEF in the first dimension, so proteins are separated according to pI and separation due to the molecular weight (Mr) in the second dimension [1]. Due to the capability a lot of possibilities were established for plant proteomic analysis. However, to obtain high quality resolution of proteins in proteomic analysis, sample preparation is important for optimal results. Plant tissue is often rich in compounds that interfere with 2-DE. Most of the difficulties in plant proteomics are associated with co-extraction of non-protein components, which affect separation of proteins and causes to protein loss [2]. Latex represents an aqueous emulsion or suspension, containing proteins, alkaloids, terpenoid structures like resins and rubber, oil starches, tannins and sugars. Due to the abundance of phenolic compounds and natural rubber particles, which interfere with 2-DE, an effective sample preparation technique is essential [3]. In our investigation we want to explore an approach to separate proteins from latices of genus Euphorbia L. in particular by 2-DE to elucidate the complex composition of containing proteins. Different sample preparation methods were tested and validate based on the number of spots that could be visualized.

Keywords: 2- DE, plant latices, sample preparation

References:

[1] O'Farell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem 1975; 250: 4007 – 4021.

[2] Wang W. Optimizing protein extraction from plant tissues for enhanced proteomics analysis. J Sep Science 2008; 31: 2032 – 2039.

[3] Lewinsohn TM. The geographical distribution of plant latex. Chemoecology 1991; 2: 64 – 68.