Selective Enrichment of Membrane Proteins by Partition Phase Separation for Proteomic Studies

[摘要] The human proteome project will demand faster, easier, and morereliable methods to isolate and purify protein targets. Membraneproteins are the most valuable group of proteins since they arethe target for 70–80% of all drugs. Perbio Science hasdeveloped a protocol for the quick, easy, and reproducibleisolation of integral membrane proteins from eukaryotic cells.This procedure utilizes a proprietary formulation to facilitatecell membrane disruption in a mild, nondenaturing environment andefficiently solubilizes membrane proteins. The technique utilizesa two-phase partitioning system that enables the class separationof hydrophobic and hydrophilic proteins. A variety of proteinmarkers were used to investigate the partitioning efficiency ofthe membrane protein extraction reagents (Mem-PER) (Mem-PER is aregistered trademark of Pierce Biotechnology, Inc) system. Theseincluded membrane proteins with one or more transmembranespanning domains as well as peripheral and cytosolic proteins.Based on densitometry analyses of our Western blots, we obtainedexcellent solubilization of membrane proteins with less than10% contamination of the hydrophobic fraction with hydrophilicproteins. Compared to other methodologies for membrane proteinsolubilization that use time-consuming protocols or expensive andcumbersome instrumentation, the Mem-PER reagents system foreukaryotic membrane protein extraction offers an easy, efficient,and reproducible method to isolate membrane proteins frommammalian and yeast cells.

[学科分类] 基础医学