Mass Spectrometry of Membrane Proteins

Abstract

Mass spectrometry as a high throughput, high speed, and high-sensitivity method has been applied widely in analyzing proteins, from identification to structural characterization. The introduction of mass spectrometry to membrane proteins analyses has accelerated the identification and functional understanding of membrane proteins. In this chapter, several MS-based approaches and applications to membrane proteins were presented, such as bottom-up and top-down proteomic, hydrogen/deuterium exchange, oxidative labeling, and native mass spectrometry. These methods have realized analyzing membrane protein at different levels, from primary structure, wherein allowing the sequence and the post-translational modifications to be determined, through to secondary structure and tertiary and quaternary structures, where the conformation dynamics and ligand-binding behavior to be defined. Finally, we provide a brief protocol of native mass spectrometry analysis of membrane proteins, which have been applied to reveal the membrane protein–protein (ligand) interaction and the conformational dynamics upon activation.