A comparative proteomic analysis of HepG2 cells incubated by S(-) and R(+) enantiomers of anti-coagulating drug warfarin.
Warfarin is a commonly prescribed oral anti-coagulant with narrow therapeutic index. It interferes with vitamin K cycle to achieve anti-coagulating effects. Warfarin has two enantiomers, S(-) and R(+) and undergoes stereoselective metabolism, with the S(-) enantiomer being more effective. We reported that the intracellular protein profile in HepG2 cells incubated with S(-) and R(+) warfarin, using iTRAQ-coupled 2-D LC-MS/MS. In samples incubated with S(-) and R(+) warfarin alone, the multi-task protein Protein SET showed significant elevation in cells incubated with S(-) warfarin but not in those incubated with R(+) warfarin. In cells incubated with individual enantiomers of warfarin in the presence of vitamin K, protein disulfide isomerase A3 which is known as a glucose-regulated protein, in cells incubated with S(-) warfarin was found to be down-regulated compared to those incubated with R(+) warfarin. In addition, Protein DJ-1 and 14-3-3 Proteinsigma were down-regulated in cells incubated with either S(-) or R(+) warfarin regardless of the presence of vitamin K. Our results indicated that Protein DJ-1 may act as an enzyme for expression of essential enzymes in vitamin K cycle. Taken together, our findings provided molecular evidence on a comprehensive protein profile on warfarin-cell interaction, which may shed new lights on future improvement of warfarin therapy.
School of Chemical and Biomedical Engineering, College of Engineering, Nanyang, Technological University, Singapore, Singapore.
Jing
J
Bai
Laleh
L
Sadrolodabaee
Chi Bun
CB
Ching
Balram
B
Chowbay
Wei
W
Ning Chen
D048948:14-3-3 Proteins; D000595:Amino Acid Sequence; D014408:Biomarkers, Tumor / Q000378:metabolism; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects*; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods; D005092:Exonucleases / Q000378:metabolism; D005095:Exoribonucleases; D005562:Formazans; D056945:Hep G2 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D007553:Isotope Labeling; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000378:metabolism; D015513:Oncogene Proteins / Q000378:metabolism; D000071617:Protein Deglycase DJ-1; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000145:classification; D020543:Proteome / Q000187:drug effects* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods; D013778:Tetrazolium Salts; D014812:Vitamin K / Q000378:metabolism; D014859:Warfarin / Q000737:chemistry / Q000494:pharmacology*
D048948:14-3-3 Proteins; D014408:Biomarkers, Tumor; D005562:Formazans; D047908:Intracellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins; D015513:Oncogene Proteins; D011506:Proteins; D020543:Proteome; D017382:Reactive Oxygen Species; D013778:Tetrazolium Salts; D014812:Vitamin K; C079782:MTT formazan; D014859:Warfarin; D005092:Exonucleases; D005095:Exoribonucleases; C413442:SFN protein, human; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1; D019704:Protein Disulfide-Isomerases; C085868:PDIA3 protein, human