Originally published In Press as doi:10.1074/mcp.M600296-MCP200 on February 11, 2007.
Molecular & Cellular Proteomics 6:953-962, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.
Research
Comparative Cytochrome P450 Proteomics in the Livers of Immunodeficient Mice Using 18O Stable Isotope Labeling*,S
Catherine S. Lane , ,
Yuqin Wang ,
Richard Betts ,
William J. Griffiths ,¶ and
Laurence H. Patterson||
From The School of Pharmacy, University of London, London WC1N 1AX, United Kingdom and the || Institute of Cancer Therapeutics, University of Bradford, West Yorkshire BD7 1DP, United Kingdom
Quantitative changes in cytochrome P450 (CYP) proteins involved in drug metabolism as a consequence of drug treatment are important parameters in predicting the fates and pharmacological consequences of xenobiotics and drugs. In this study we undertook comparative P450 proteomics using liver from control and 1,4-bis-2-(3,5-dichloropyridyloxybenzene) (TCPOBOP)-dosed mice. The method involved separation of microsomal proteins by SDS-PAGE, trypsin digestion, and postdigest 18O/16O labeling followed by nano-LC-MS/MS for peptide identification and LC-MS for relative quantification. Seventeen P450 proteins were identified from mouse liver of which 16 yielded data sufficient for relative quantification. All the P450s detected were unambiguously identified except the highly homologous CYP2A4/2A5. With the exception of CYP2A12, -2D10, and -2F2, the levels of all the P450s quantified were affected by treatment with TCPOBOP (3 mg/kg). CYP1A2, -2A4/5, -2B10, -2B20, -2C29, -2C37, -2C38, -3A11, and -39A1 were up-regulated, and CYP2C40, -2E1, -3A41, and -27A1 down-regulated. The response of CYP2B20 to stimulation has not been distinguished previously from that of CYP2B10 because of the poor discrimination between these two proteins (they share 87% sequence identity). Differential response to chemical stimulation by closely related members of the CYP2C subfamily was also observed.
¶ To whom correspondence should be addressed. Tel.: 44-207753-5876; Fax: 44-207753-5964; E-mail: william.griffiths{at}pharmacy.ac.uk

CiteULike Complore Connotea Del.icio.us Digg Reddit Technorati What's this?
This article has been cited by other articles:

|
 |

|
 |
 
X. Ye, B. Luke, T. Andresson, and J. Blonder
18O Stable Isotope Labeling in MS-based Proteomics
Briefings in Functional Genomics,
March 1, 2009;
8(2):
136 - 144.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A.-M. Yu, J. Qu, M. A. Felmlee, J. Cao, and X.-L. Jiang
Quantitation of Human Cytochrome P450 2D6 Protein with Immunoblot and Mass Spectrometry Analysis
Drug Metab. Dispos.,
January 1, 2009;
37(1):
170 - 177.
[Abstract]
[Full Text]
[PDF]
|
 |
|
Copyright © 2007 by the American Society for Biochemistry and Molecular Biology.
|
Advertisement
Advertisement
|