Proteomic response analysis of endothelial cells of human coronary artery to stimulation with carbachol.
OBJECTIVE
To identify the molecular basis of the endothelial target for acetylcholine (ETA).


METHODS
Proteomic methods were used to monitor changes in protein expression in the first 10 h following the stimulation of human coronary endothelial cells with carbachol 100 micromol/L. Thirty proteins showing the largest changes were identified by mass spectrometry.


RESULTS
Based on analysis with Image Master 2-D Elite software, about 623 protein spots were detected in control cells and 825 protein spots in carbachol-treated cells, the matching rate was 68.1 %. Among all the detected spots, 39 were up-regulated and 29 were down-regulated, showing detectable changes varied from 1.7-3.8 folds. Forty one spots in the peptide mass fingerprints were successfully obtained. The most interesting feature was that all the four newly synthesized proteins belonged to the heat shock protein family.


CONCLUSIONS
These identified proteins played key roles in the molecular mechanism of ETA.
Yu
Min
M
Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China.
Chen
Dong-Mei
DM
Hu
Gang
G
Wang
Hai
H
D000109:Acetylcholine / Q000031:analogs & derivatives; D002217:Carbachol / Q000494:pharmacology*; D018679:Cholinergic Agonists / Q000494:pharmacology*; D003331:Coronary Vessels / Q000166:cytology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D042783:Endothelial Cells / Q000737:chemistry*; D006360:Heat-Shock Proteins / Q000032:analysis; D006801:Humans; D011506:Proteins / Q000032:analysis*; D040901:Proteomics
D018679:Cholinergic Agonists; D006360:Heat-Shock Proteins; D011506:Proteins; D002217:Carbachol; D000109:Acetylcholine