Professors Morteza G. Khaledi and David C. Muddiman of North Carolina State University are supported by the Chemical Measurement and Imaging (CMI) Program in the Division of Chemistry to investigate the use of newly discovered supramolecular liquids called Perfluoro Alcohol Induced Coacervates (PFAIC) for enhanced selective extraction and enrichment of membrane proteins from biological matrices. Membrane proteins are the most important cell surface membrane constituents. They act as highly-selective cellular receptors and gatekeepers, regulating the traffic of critical nutrients and energy in and out of the cells. Solubilization, extraction, and enrichment of membrane proteins from biological samples into aqueous solutions suitable for further analysis - a critical step for the characterization of the individual proteins and for understanding their cellular function - is an area of considerable challenge in contemporary bioanalytical chemistry and molecular biology. The project promises to tackle this challenge and provide new avenues for the identification of membrane proteins and hence advance our understanding of cellular function and life processes. In the course of conducting the project, graduate and undergraduate students will gain valuable skills in bioanalytical separation technologies, proteomics and mass spectrometry. Plans are also established, through a partnership with the Kenan Fellows Program of the Kenan Institute at North Carolina State University, to involve high school teachers in summer research and in the development of high school curriculum materials in the general areas related to this research.
The overarching objective of the proposed research is to explore the use of Perfluoro Alcohol Induced Coacervates (PFAIC) for enhanced selective extraction and enrichment of membrane proteins from biological matrices. The proposed research is organized into four specific aims: (1) to achieve an understanding of the principles of extraction of proteins in PFAIC systems through systematic studies of partitioning patterns, extraction efficiency and enrichment factors of proteins with different structural and physico-chemical properties and PFAIC systems with different chemical composition; (2) to investigate general strategies for enrichment of membrane proteins into PFAIC phases; (3) to investigate the effectiveness of PFAIC for pre-fractionation, extraction and enrichment of membrane proteins in shot-gun bottom-up proteomics analysis of yeast as a model biological sample; and (4) to explore the possibility of inducing coacervation of lipid bilayers in cell membranes and concomitant solubilization of the membrane bound proteins.
