Two-dimensional electrophoretic separation of subproteomes in adult rat hippocampal subregions after membrane protein enrichment with sequential extraction
Abstract：Objective To establish a method for analysis of the subproteomes following membrane protein enrichment, so as to investigate the differences in cytosolic and membrane protein/peptide contents among different hippocampal subregions (CA1,CA3 and dentate gyrus) of rats. Methods The hippocampal subregions were isolated by microdissection, and after membrane protein enrichment with protein solubility-based sequential sample extraction, subproteome profiling was accomplished using
modified two-dimensional electrophoresis and silver staining method compatible with high-performance mass spectrometry,which displayed cytosolic or membrane proteins/peptides separately. The efficacy of the electrophoresis was evaluated by image analyzing software. Results The membrane proteins were enriched by 8 folds, and around 30％ more spots were shown
on the integrated dentate gyrus map than on the conventional map obtained by one-step protein extraction. Such improvement could also be seen in protein mapping of CA1 and CA3 regions. Conclusion When analyzing hippocampal subregions sequential extraction and two-dimensional electrophoresis more effectively separate and display enriched membrane protein/peptides, which may play important roles in such cellular events as signal transduction.
Keywords：proteomicshippocampusmicrodissectionsequential extractionmembrane proteins
Publication Date：2004-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 1217-1222,1226 )