[Two-dimensional electrophoretic separation of subproteomes in adult rat hippocampal subregions after membrane protein enrichment with sequential extraction].
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.


CONCLUSIONS
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.
Chen
Liang
L
Department of Physiology, Southern Medical University, Guangzhou 510515, China.
Yang
Jian-ming
JM
Li
Xiao-ming
XM
Li
Xiao-wen
XW
Gao
Tian-ming
TM
D000818:Animals; D018891:Dentate Gyrus / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D006624:Hippocampus / Q000737:chemistry*; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification*; D042282:Microdissection; D010449:Peptide Mapping; D020543:Proteome / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar
D008565:Membrane Proteins; D020543:Proteome