Cytochrome P450 expression

Uno, T., Nakao, A., Masuda, S. et al. (2006) Modification of small molecules by using cytochrome P450 expressed in Escherichia coli. Journal of Industrial Microbiology Biotechnology, 33, 1043-1050. 

Ioannides C (2006) Cytochrome P450 expression in the liver of food-producing animals. Curr. Drug Metab. 7 335-348. 

Vondracek M, Xi Z, Larson P, Baker V, Mace K, Pfeifer A, Tjalve H, Donato MT, Gomez-Lechon MJ, and Grafstrom C (2001) Cytochrome P450 expression and related metabolism in human buccal mucosa. Carcinogenesis 22 481-488. 

Blume N, Leonard J, Xu ZJ, Watanabe O, Remotti H, et al. 2000. Characterization of Cyp2d22, a novel cytochrome P450 expressed in mouse mammary cells. Arch Biochem Biophys 381 191-204. 

Warner M, Ahlgren R, Zaphiropoulos PG, Hayashi S, Gustafsson JA. 1991. Identification and localization of cytochromes P450 expressed in brain. Methods Enzymol 206 ... 

Madan A, Graham RA, CarroU KM, Mudra DR, Burton LA, Krueger LA, Downey AD, Czerwinski M, Forster J, Ribadeneira MD, Gan LS, LeCluyse EL, Zech K, Robertson P, Jr., Koch P, Antonian L, Wagner G, Yu L, Parkinson A (2003) Effects of prototypical microsomal enzyme inducers on cytochrome P450 expression in cultured human hepatocytes. Drug Metab Dispos31(4) 421 31... 

It is of interest that studies with cloned human cytochromes P450 expressed in insect SF9 cells have shown that only CYP2A6 catalysed 7-hydroxylation, while the formation of orl/20-hydroxyphenylacetaldehyde was supported by CYPlAl, 1A2, 2B6, 2E1 and 3A4 (Zhuo et al, 1999). 

Use the system with the highest available expression levels. This will at the very least simplify analytical procedures and shorten incubation times. Levels of cytochrome P450 expression which can be quantitated spectrophotometrically allow more accurate quantitation of expression levels for comparisons among different P450 forms (on a nmol enzyme basis). 

Use a system which allows the preparation of subcellular fractions. Microsome preparation provides a 5- to 10-fold enrichment in cytochrome P450 expression levels and also permits longer incubation times. This enrichment and concomitant increase in metabolizing capacity per unit volume may be important in detecting metabolism of slow turnover substrates. 

The first transient expression system for cytochrome P450 expression was the COS cell system which couples monkey cells with an SV40-based expression vector (Zuber et al., 1986). The cDNA of interest is introduced into a plasmid expression vector under control of a heterologous promoter. The vector is introduced into COS cells via electroporation or another method allowing DNA uptake. The plasmid vector replicates within the cells and cDNA-derived protein accumulates within the cell. 

Kato R, Yamozoe Y. Sex-dependent regulation of cytochrome P450 expression. In Ruckpaul K, Rein H, eds. Frontiers of Bio transformation, Vol. II, Principles, Mechanisms and Biological Consequences of Induction. London Taylor Francis, 1990. 

Vondracek, M., et al. 2001. Cytochrome P450 expression and related metabolism in human buccal mucosa. Carcinogenesis 22 481. 

Gonzalez FJ, Korzekwa KR. Cytochromes P450 expression systems. Annu Rev Pharmacol Toxicol 1995 35 369-390. 

LeCluyse EL. Human hepatocyte culture systems for the in vitro evaluation of cytochrome P450 expression and regulation. Eur J Pharm Sci 2001 13(4) 343—368. 

Madan A, Graham RA, Carroll KM, et al. Effects of prototypical microsomal enzyme inducers on cytochrome P450 expression in cultured human hepatocytes. Drug Metab Dispos 2003 31 421—431. 

Treluyer JM, Cheron G, Sonnier M, Cresteil T (1996) Cytochrome P450 expression in sudden infant death syndrome. Biochem Pharmacol, 52 497-504. 

Hung, C.F., Harrison, T.L., Berenbaum, M.R. and Schuler, M.A. (1995) CYP6B3 a second furanocoumarin-inducible cytochrome P450 expressed in Papilio polyxenes. Insect Mol. Biol., 4, 149-60. 

Lechevrel M, Casson AG, Wolf CR, Hardie LJ, Flinterman MB, et al. 1999. Characterization of cytochrome P450 expression in human oesophageal mucosa. Carcinogenesis 20 243 48... 

Miranda, C.L., P. Collodi, X. Zhao, D.W. Barnes and D.R. Buhler. Regulation of cytochrome P450 expression in a novel liver cell line from zebrafish (Brachydanio rerio). Arch. Biochem. Biophys. 305 320-327, 1993. 

Belloc, C., Baird, S., Cosme, J., Lecoeur, S., Gautier, J.-C., Challine, D., de Waziers, I., Flinois J.-P., and Beaune, P.H. (1996) Human cytochrome P450 expressed in Escherichia coli production of spedfic antibodies. Toxicology, 106, 207-219. 

K. Ohyama, N. Hatanaka, S. Asahi et al. (2002). Roles of NADPH-P450 reductase and apo- and holo-cytochrome b5 on xenobiotic oxidations catalyzed by 12 recombinant human cytochrome P450s expressed in membranes of Escherichia coli. Protein Expr. Purif. 24, 329-337. 

Middle-age alterations in the sexually dimorphic plasma growth hormone profiles Involvement of growth hormone-releasing factor and effects on cytochrome p450 expression. Drug Melab. Dispos. 30, 141-147. 

Henderson, C.J. and C.R. Wolf (1991). Evidence that the androgen receptor mediates sexual differentiation of mouse renal cytochrome P450 expression. Biochem. J. 278, 499-503. 

Swanson HI (2004) Cytochrome P450 expression in human keratinocytes an aryl hydrocarbon receptor perspective. Chem Biol Interact 149 69-79 Uetrecht J (2001) Prediction of a new drag s potential to cause idiosyncratic reactions. Curr Opin Drug Discov Devel 4 55-59... 

---