Cytochrome fusion protein

Rodrigues, A.D. Mulford, D.J. Lee, R.D. Surber, B.W. Kukulka, M.J. Ferrero, J.L. Thomas, S.B. Shet, M.S. Estabrook, R.W. In vitro metabolism of terfenadine by a purified recombinant fusion protein containing cytochrome P4503A4 and NADPH-P450 reductase. Comparison to human liver microsomes and precision-cut liver tissue slices. Drug Metab.Dispos., 1995, 23, 765-775... 

Fisher, C.W., M.S. Shet, D.L. Caudle, C.A. Martin-Wixtrom, and R.W. Estabrook (1992). High-level expression in Escherichia coli of enzymatically active fusion proteins containing the domains of mammalian cytochromes P450 and NADPH-P450 reductase flavoprotein. Proc. Natl. Acad. Sci. USA 89, 10817-10821. 

Chun, Y.J., X. Shimada, and F.P. Guengerich (1996). Construction of a human cytochrome P450 lAl Rat NADPH-cytochrome P450 reductase fusion protein cDNA and expression in Escherichia coli, purification, and catalytic properties of the enzyme in baeterial eells and after purification. Arch. Biochem. Biophys. 330,48—58. 

Deeni, Y.Y., M.J. Paine, A.D. Ayrton, S.E. Clarke, R. Chenery, and C.R. Wolf (2001). Expression, purification, and biochemical characterization of a human cytochrome P450 CYP2D6-NADPH cytochrome P450 reductase fusion protein. Arch. Biochem. Biophys. 396, 16-24. 

Sibbesen, O., J.J. De Voss, and PR. Ortiz de Montellano (1996). Putidaredoxin reductase-putidaredoxin-cytochrome P450cam triple fusion protein. Construction of a self-sufficient Escherichia coli catalytic system. J. Biol. Chem. 271, 22462-22469. 

Jackson, C.J., D.C. Lamb, T.H. Marczylo, A.G. Warrilow, N.J. Manning, D.J. Lowe et al. (2002). A novel sterol 14alpha-demethylase/ferredoxin fusion protein (MCCYP51FX) from Methylo-coccus capsulatus represents a new class of the cytochrome P450 superfamily. J. Biol. Chem. 277, 46959 6965. 

Shet, M.S., C.W. Fisher, PL. Holmans, and R.W. Estabrook (1993). Human cytochrome P450 3A4 Enzymatic properties of a purified recombinant fusion protein containing NADPH-P450 reductase. Proc. Natl Acad. Scl USA 90, 11748-11752. 

Figure 13.2. A proposal for a simplified classification of CYPs with reference to either use of a ferredoxin or alternative electron transport mechanism. Class la— typical bacterial system supported by ferredoxin and ferredoxin reductase, for example, CYPlOl Class Ib—Rhodococcus sp. CYP fusion protein containing a ferredoxin domain Class Ic—Methylococcus capsulatus CYPS 1-ferredoxin fusion . Class 11a— typical eukaryotic CYP/NADPH-cytochrome P450 reductase system Class lib—a fusion protein of a CYP and flavoprotein reductase, for example, P450gM 3 Class IIc—P450 j containing separate flavodoxin and flavodoxin reductase partners . Class fll—standalone functional CYPs, for example, P450jj. ...

In the biological chemistry field, fusion proteins have recently shown more potential as organelle-specific labels. For subcellular analysis, fusion proteins can be expressed that contain both a fluorescent protein and a subcellular localization sequence. This combination results in organelle-specific labeling that retains the excellent photochemical properties of fluorescent proteins (e.g., intense fluorescence and photostability). We commonly use a commercially available plasmid that codes for a fusion protein of red fluorescent protein (DsRed2) and the mitochondrial targeting sequence from subunit VIII of cytochrome c oxidase, which contains the neomycin/kanamycin resistance gene, as described below. 

Brodhun F, Gobel C, Hornung E, Feussner I (2009) Identification of PpoA from Aspergillus nidulans as a fusion protein of a fatty acid heme dioxygenase/ peroxidase and a cytochrome P450. J Biol Chem 284 11792-11805... 

Lamb DC, Kelly DE, Hanley SZ, Mehmood Z, Kelly SL (1998) Glyphosate is an inhibitor of plant cytochrome P450 functional expression of Thlaspi arvensae cytochrome P45071Bl/reduetase fusion protein in Escherichia coli. Bioehem Biophys Res Commun 244 110 114... 

Hotze M, Schroder G, Schrodo- J (1995) Cinnamate 4-hydroxylase fiom Catharanthusroseus, and a strategy for the functional expression of plant cytochrome P450 proteins as translational fusions with P450 reductase in Escherichia coli. FEES Lett 374 345-350... 

Kim DH, Kim B-G, Jung NR, Ahn J-H (2(X)9) Production of genistein from naringenin using Escherichia coli containing isoflavone synthase-cytochrome P450 reductase fusion protein. J Microbiol Biotechnol 19 1612-1616... 

For example, the N-terminal human CYP2D6 sequence was linked to the C-terminal human cytochrome reductase module, resulting in a self-sufficient enzyme exhibiting activity toward a wide range of pharmaceutical compounds such as bufuralol, metoprolol, or dextromethorphan [152]. Other examples are fusion proteins ofhumanCYPlAl with rat CPR [153], human CYP2C9 with BMR [154], and... 

---