Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
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Bioactivation of Aromatic Amines by Human CYP2W1, An Orphan Cytochrome P450 Enzyme

  • Received : 2010.07.29
  • Accepted : 2010.08.14
  • Published : 2010.09.01
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Abstract

The human genome contains approximately 13 orphan cytochrome P450 (P450, CYP) genes, of which the apparent function or substrate has not been identified. However, they seem to possess their own biological relevance in some tissues or developmental stages. Here, we characterized the heterologously expressed CYP2W1, an orphan P450 enzyme. The recombinant CYP2W1 protein containing a $6{\times}$(His)-tag at Nterminus has been expressed in Escherichia coli and purified. Expression level of CYP2W1 holoenzyme was around 500 nmol P450 holoenzyme per liter culture medium. The reduced CO difference spectrum of CYP2W1 showed a maximum absorption at 449 nm. CYP2W1 indicated the significant induction to bioactivate Trp-P-1, MeIQ, and IQ in E. coli DJ701 tester strain. However, the bioactivation of B[$\alpha$]P, and NNK by CYP2W1 was relatively low. The model structure of CYP2W1 suggested the characteristic P450 folds with the lengths and orientations of the individual secondary elements. The F-G loop is situated on the distal side of heme to accommodate the flexibility of active site of CYP2W1. These studies can provide useful information for the finding of its biological roles and structure-function relationships of an orphan CYP2W1 enzyme.

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References

  1. Arnold, K., Bordoli, L., Kopp, J. and Schwede, T. (2006). The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics, 22, 195-201. https://doi.org/10.1093/bioinformatics/bti770
  2. Choudhary, D., Jansson, I., Stoilov, I., Sarfarazi, M. and Schenkman, J.B. (2005). Expression patterns of mouse and human CYP orthologs (families 1-4) during development and in different adult tissues. Arch. Biochem. Biophys., 436, 50-61. https://doi.org/10.1016/j.abb.2005.02.001
  3. Gomez, A., Karlgren, M., Edler, D., Bernal, M.L., Mkrtchian, S. and Ingelman-Sundberg, M. (2007). Expression of CYP2W1 in colon tumors: regulation by gene methylation. Pharmacogenomics, 8, 1315-1325. https://doi.org/10.2217/14622416.8.10.1315
  4. Guengerich, F.P. (1993). Cytochrome P450 enzymes. American Scientist, 81, 440-447.
  5. Guengerich, F.P. (2008). Cytochrome p450 and chemical toxicology. Chem. Res. Toxicol., 21, 70-83. https://doi.org/10.1021/tx700079z
  6. Guengerich, F.P., Tang, Z., Cheng, Q. and Salamanca-Pinzon, S.G. (2010a). Approaches to deorphanization of human and microbial cytochrome P450 enzymes. Biochim. Biophys. Acta., (in press).
  7. Guengerich, F.P., Tang, Z., Salamanca-Pinzon, S.G. and Cheng, Q. (2010b). Characterizing proteins of unknown function: orphan cytochrome p450 enzymes as a paradigm. Mol. Interv., 10, 153-163. https://doi.org/10.1124/mi.10.3.6
  8. Guengerich, F.P., Wu, Z.L. and Bartleson, C.J. (2005). Function of human cytochrome P450s: characterization of the orphans. Biochem. Biophys. Res. Commun., 338, 465-469. https://doi.org/10.1016/j.bbrc.2005.08.079
  9. Josephy, P.D. (2000). The Escherichia coli lacZ reversion mutagenicity assay. Mutat. Res., 455, 71-80. https://doi.org/10.1016/S0027-5107(00)00063-4
  10. Karlgren, M., Gomez, A., Stark, K., Svard, J., Rodriguez-Antona, C., Oliw, E., Bernal, M.L., Ramon y Cajal, S., Johansson, I. and Ingelman-Sundberg, M. (2006). Tumor-specific expression of the novel cytochrome P450 enzyme, CYP2W1. Biochem. Biophys. Res. Commun., 341, 451-458. https://doi.org/10.1016/j.bbrc.2005.12.200
  11. Kim, D. and Guengerich, F.P. (2005). Cytochrome P450 activation of arylamines and heterocyclic amines. Annu. Rev. Pharmacol. Toxicol., 45, 27-49. https://doi.org/10.1146/annurev.pharmtox.45.120403.100010
  12. Kim, D., Wu, Z.L. and Guengerich, F.P. (2005). Analysis of coumarin 7-hydroxylation activity of cytochrome P450 2A6 using random mutagenesis. J. Biol. Chem., 280, 40319-40327. https://doi.org/10.1074/jbc.M508171200
  13. Li, W., Tang, Y., Hoshino, T. and Neya, S. (2009). Molecular modeling of human cytochrome P450 2W1 and its interactions with substrates. J. Mol. Graph. Model, 28, 170-176. https://doi.org/10.1016/j.jmgm.2009.06.002
  14. Nelson, D.R., Zeldin, D.C., Hoffman, S.M., Maltais, L.J., Wain, H.M. and Nebert, D.W. (2004). Comparison of cytochrome P450 (CYP) genes from the mouse and human genomes, including nomenclature recommendations for genes, pseudogenes and alternative-splice variants. Pharmacogenetics, 14, 1-18. https://doi.org/10.1097/00008571-200401000-00001
  15. Omura, T. and Sato, R. (1964). The Carbon Monoxide-Binding Pigment of Liver Microsomes. I. Evidence for Its Hemoprotein Nature. J. Biol. Chem., 239, 2370-2378.
  16. Rowland, P., Blaney, F.E., Smyth, M.G., Jones, J.J., Leydon, V.R., Oxbrow, A.K., Lewis, C.J., Tennant, M.G., Modi, S., Eggleston, D.S., Chenery, R.J. and Bridges, A.M. (2006). Crystal structure of human cytochrome P450 2D6. J. Biol. Chem., 281, 7614-7622. https://doi.org/10.1074/jbc.M511232200
  17. Tang, Z., Martin, M.V. and Guengerich, F.P. (2009). Elucidation of functions of human cytochrome P450 enzymes: identification of endogenous substrates in tissue extracts using metabolomic and isotopic labeling approaches. Anal. Chem., 81, 3071-3078. https://doi.org/10.1021/ac900021a
  18. Wu, Z.L., Sohl, C.D., Shimada, T. and Guengerich, F.P. (2006). Recombinant enzymes overexpressed in bacteria show broad catalytic specificity of human cytochrome P450 2W1 and limited activity of human cytochrome P450 2S1. Mol. Pharmacol., 69, 2007-2014. https://doi.org/10.1124/mol.106.023648
  19. Yun, C.-H., Miller, G.P. and Guengerich, F.P. (2000). Rate-determining steps in phenacetin oxidations by human cytochrome P450 1A2 and selected mutants. Biochemistry, 39, 11319-11329. https://doi.org/10.1021/bi000869u
  20. Zhou, H., Josephy, P.D., Kim, D. and Guengerich, F.P. (2004). Functional characterization of four allelic variants of human cytochrome P450 1A2. Arch. Biochem. Biophys., 422, 23-30. https://doi.org/10.1016/j.abb.2003.11.019

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