title: Rosuvastatin-regulated post-translational phosphoproteome in human umbilical vein endothelial cells.
abstract: Statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, are widely prescribed as cholesterol-lowering drugs. Statins have recently been found to have pleiotropic effects that are independent of their lipid-lowering properties. Phosphorylation of serine, threonine, and tyrosine residues of functional proteins are considered to be important in the endothelial signaling cascade. In this study, protein phosphorylation status in human umbilical vein endothelial cells (ECs) after rosuvastatin treatment was examined. The proteins were collected from rosuvastatin-treated ECs and then the phosphorylated peptides purified by a Fe(3+)-immobilized metal-affinity chromatography bead system were examined by liquid chromatography-tandem mass spectrometry analysis. Alterations of the phosphorylation status of proteins were noticed after rosuvastatin treatment. There were 277 and 530 phosphorylated proteins identified from the control and rosuvastatin-treated ECs, respectively. Among those proteins, T78, in addition to S156 of the Ras-GTPase-activating protein, was phosphorylated after rosuvastatin treatment. Rosuvastatin reduced the phosphorylation of Y455 in HSP90 protein. Decreased phosphorylation of T211 with a concurrent increase in the T291 phosphorylation of Akt1 was observed under rosuvastatin treatment. Increased S633 phosphorylation was detected in endothelial nitric oxide synthase. Western blot analysis further showed an earlier and greater S633 phosphorylation than that of S1177 in endothelial nitric oxide synthase after rosuvastatin treatment. Changes in the phosphorylation status of these proteins may alter the protein's function and affect endothelial physiology. The current study provides new insights leading to a better understanding of the pleiotropic effects of statins on the vascular system.
authors.affiliation: Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Bin
authors.initials: B
authors.lastname: Huang
authors.forename: Chien-Hsing
authors.initials: CH
authors.lastname: Wu
authors.forename: Fu-An
authors.initials: FA
authors.lastname: Li
authors.forename: Shih-Shin
authors.initials: SS
authors.lastname: Liang
authors.forename: Ying-Hua
authors.initials: YH
authors.lastname: Shieh
authors.forename: Ling Danny
authors.initials: LD
authors.lastname: Wang
mesh_terms: D020816:Amino Acid Motifs; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D004265:DNA Helicases; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005464:Fluorobenzenes / Q000494:pharmacology*; D005786:Gene Expression Regulation; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D008969:Molecular Sequence Data; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D000075223:Poly-ADP-Ribose Binding Proteins; D020543:Proteome / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology*; D020365:RNA Helicases; D000067776:RNA Recognition Motif Proteins; D000068718:Rosuvastatin Calcium; D015398:Signal Transduction; D013449:Sulfonamides / Q000494:pharmacology*
chemical_list: D002352:Carrier Proteins; D005464:Fluorobenzenes; D018841:HSP90 Heat-Shock Proteins; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D010750:Phosphoproteins; D000075223:Poly-ADP-Ribose Binding Proteins; D020543:Proteome; D011743:Pyrimidines; D000067776:RNA Recognition Motif Proteins; D013449:Sulfonamides; D000068718:Rosuvastatin Calcium; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; C494918:AKT1 protein, human; D051057:Proto-Oncogene Proteins c-akt; D004265:DNA Helicases; C116070:G3BP1 protein, human; D020365:RNA Helicases