Comparison of Multidimensional Separation Strategies for Studying Membrane Proteomes

摘要

Membrane proteins account for more than one third of open reading frames in the human genome and play crucial roles in many biological processes. Scores of examples have shown a direct link between the disregulation of membrane protein biology and human diseases. Membrane proteins also represent a large number of known drug targets. However, this class of proteins remains an under-represented subset of proteins accessible to mass spectrometric analysis, due in part to their inherent amphipathic nature. Here we demonstrate a new strategy for membrane protein analysis, by incorporating a recently developed high-recovery macroporous reversed-phase (RP) C18 (mRP-C18) column separation (1) with additional in-gel or in-solution fractionation to further reduce sample complexity prior to microfluidic chip-based nano-LC/MS analysis. In this study, membrane preparations of HeLa cells were used to demonstrate the feasibility of this approach (Figure 1).