Rosuvastatin Modulates the Post-Translational Acetylome in Endothelial Cells.
BACKGROUND
Statins are lipid-lowering drugs that can simultaneously evoke pleiotropic effects on cardioprotection, vasodilation, and diabetes prevention. Recently, statins have been reported to be able to activate the AMP-activated protein kinase, thereby up-regulating sirtuin (SIRT) that functions as non-histone deacetylases. Therefore, it is essential to investigate the post-translational acetylome that might explain the mechanism of statin-modulated pleiotropic effects.


METHODS
Endothelial cells EAhy 926 treated with rosuvastatin were used to monitor the expression of SIRTs proteins. The protein lysates of both mock- and rosuvastatin-treated cells were further separated by two- dimensional gel electrophoresis coupled with western blotting analysis. The significantly changed acetyl- containing proteins detected by using an anti-acetyl lysine antibody were collected from another preparative gel for mass spectrometric assay to identify the acetylated site in the proteins.


RESULTS
Rosuvastatin treatment was shown to increase the SIRT1 expression when compared with SIRT2. Among 100 detected proteins with acetylated signal, 12 showed an increased level of acetylation, whereas 6 showed a decreased level of acetylation (deacetylation). The acetylated lysine (K) sites of 3 heat shock proteins, i.e., HSP47/K(165), HSP70/K(380), and heat shock-inducible protein/K(417), were determined. We also found that beta-filamin, elongation factor, galectin and hCG22067 have 2 acetylated lysine sites in their peptide sequences. These dynamic acetylations might alter the protein's function and are thought to be important in regulating statin-mediated pleiotropic effect.


CONCLUSIONS
Our study provided a feasible methodology for detecting acetylated proteins. This acetylome information may be utilized to explain, at least partially, the mechanisms of statin-derived pleiotropic effects.


BACKGROUND
Acetylation/deacetylation; Acetylome; Endothelial cell; Proteomics; Rosuvastatin; Sirtuin.
Department of Anesthesiology, Chi Mei Medical Center, Tainan, Taiwan; ; Department of Medical Laboratory Science and Biotechnology, Chung Hwa University of Medical Technology, Tainan, Taiwan;
Ming Chung
MC
Lin
Department of Anesthesiology, Chi Mei Medical Center, Tainan, Taiwan;
Chung Hsi
CH
Hsing
Institute of Biomedical Science, Academia Sinica, Taipei, Taiwan;
Fu An
FA
Li
Division of Nephrology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan;
Chien Hsing
CH
Wu
Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung, Taiwan;
Yaw Syan
YS
Fu
Department of Anesthesiology, Mackay Memorial Hospital, Taipei, Taiwan.
Jen Kun
JK
Cheng
Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung, Taiwan;
Bin
B
Huang