Identification and characterization of membrane proteins from mouse brain tissue

Abstract

Membrane proteins are attractive targets for proteomics research because of their vital roles in numerous processes of the cell including: cell adhesion, immune response, metabolism and signal transduction... At the start of proteomics research, two-dimensional gel electrophoresis (2D-PAGE) was routinely used to separate complex proteomic samples. However, this method faces difficulties in separating membrane proteins due to their hydrophobicity. In this study, mouse brain membrane fractions were prepared using carbonate extraction and ultracentrifugation. The separation and identification of the membrane proteins by using a gel-based approach in combination with two-dimensional nano liquid chromatography coupled online with tandem mass spectrometry (2DNanoLC-Q-TOF-MS/MS) were presented. In total, 298 identified membrane proteins from mouse brain tissues were verified and predicted by UniProt database, SOSUI and TMHMM algorithms. Among them, 129 (43.3%) proteins that have at least one transmembrane domain were predicted by SOSUI and TMHMM. Furthermore, the function, subcellular location and hydrophobicity value of the identified membrane proteins were also categorized. © 2013 Dung NT, et al.