A proteomic method for analysis of CYP450s protein expression changes in carbon tetrachloride induced male rat liver microsomes.
Carbon tetrachloride (CCl(4)) is a well-known model compound for producing chemical hepatic injury. Cytochrome P450 is an important monooxygenase in biology. We investigated the CYP450 protein expression in the in vivo hepatotoxicity of rats induced by CCl(4). In this experiment, CCl(4) were administered to male rats, and their livers at 24h post-dosing were applied to the proteomic analysis. Blood biochemistry and histopathology were examined to identify specific changes. At the same time, a novel acetylation stable isotopic labeling method coupled with LTQ-FTICR mass spectrometry was applied to disclose the changes of cytochrome P450 expression amounts. The quantitative proteomics method demonstrated its correlation coefficient was 0.9998 in a 100-fold dynamic range and the average ratio of the labeled peptides was 1.04, which was very close to the theoretical ratio of 1.00 and the standard deviation (S.D.) of 0.21. With this approach, 17 cytochrome P450 proteins were identified and quantified with high confidence. Among them, the expression amount of 2C11, 3A2, and 2 E1 were down-regulated, while that of 2C6, 2B2, and 2B1 were up-regulated.
Jia
Nuan
N
Shanghai Key Laboratory for Pharmaceutical Metabolites Research, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China.
Liu
Xin
X
Department of Genomics and Proteomics, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing 100850, PR China.
Wen
Jun
J
Shanghai Key Laboratory for Pharmaceutical Metabolites Research, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China.
Qian
Linyi
L
Department of Genomics and Proteomics, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing 100850, PR China.
Qian
Xiaohong
X
Department of Genomics and Proteomics, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing 100850, PR China.
Wu
Yutian
Y
Shanghai Key Laboratory for Pharmaceutical Metabolites Research, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China.
Fan
Guorong
G
Shanghai Key Laboratory for Pharmaceutical Metabolites Research, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China. Electronic address: guorfan@yahoo.com.cn.
D000345:Affinity Labels; D000595:Amino Acid Sequence; D000818:Animals; D002251:Carbon Tetrachloride / Q000633:toxicity*; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000097:blood; D004591:Electrophoresis, Polyacrylamide Gel; D007527:Isoenzymes; D007553:Isotope Labeling; D008099:Liver / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D008297:Male; D013058:Mass Spectrometry; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D008969:Molecular Sequence Data; D009336:Necrosis; D010452:Peptide Biosynthesis; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards
D000345:Affinity Labels; D007527:Isoenzymes; D003577:Cytochrome P-450 Enzyme System; D002251:Carbon Tetrachloride