Coacervation is a form of organized self-assembly where amphiphilic molecules aggregate and form two separate phases in aqueous-based media. The two-phase system consists of an amphiphile-rich phase called coacervate which is in equilibrium with a second aqueous-rich phase containing a small level of amphiphile. Formation of the two-phase system can be induced by perfluoro alcohols and acids under various conditions. The combination of an amphiphile, tetra-<i>n</i>-butylammonium bromide (TBA), and a fluorinated alcohol, tetrafluoroethylene (TFE), under various pH and/or ion concentrations, were studied. The coacervate systems were used to extract model hydrophilic and hydrophobic proteins. The systems of interest for membrane proteomic applications are those that show higher enrichment and selectivity towards hydrophobic proteins (higher grand average of hydropathy). Of the systems observed, TBA /TFE showed a higher membrane protein selectivity in the coacervate phase. This system was utilized to extract and enrich proteins from baker’s yeast, <i>Saccharomyces cerevisiae</i>, a well-defined organism in the study of proteomics. A mixture containing TBA/TFE and the crude yeast lysate was vortexed and centrifuged. The two phases formed were separated and filter aided sample preparation was applied to individual phases prior to downstream proteomic analysis by liquid chromatography-mass spectrometry. An approximately 24% increase in intrinsic membrane protein identification was observed. In addition, more hydrophobic proteins were identified in the coacervate phase. Such results can be applied in the diagnostic study of gene mutation diseases, such as breast cancer, where the overexpression of a membrane protein is a key observation.

Fluoroalcohol-induced coacervation in membrane proteomics

255th National Meeting (2018)

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