Cytochrome recombinant

Yamazaki, H., Ueng, Y.F., Shimada, T. and Guengerich, F.P. (1995) Roles of divalent metal ions in oxidations catalyzed by recombinant cytochrome P450 3A4 and replacement of NADPH-cytochrome P450 reductase with other flavoproteins, ferredoxin, and oxygen surrogates. Biochemistry, 34, 8380—8389. 

HALKIER, B.A., NIELSEN, H.L., KOCH, B., M0LLER, B.L., Purification and characterization of recombinant cytochrome P450TYR expressed at high levels in Escherichia coli, Arch. Biochem. Biophys., 1995,322, 369-377. 

BARNES, H.J., ARLOTTO, M.P., WATERMAN, M.R., Expression and enzymatic activity of recombinant cytochrome P450 17a-hydroxylase in Escherichia coli, Proc. Natl. Acad. Sci. USA, 1991,88, 5597-5601. 

Cerep evaluates hits, leads, and new compounds from the metabolic point of view using liver microsomes and recombinant cytochrome P450s (275). The results of these metabolic screening studies offer insight to the rate of metabolic pathways, in vivo pharmacokinetic properties, and drug-drug interactions. 

Recombinant Cytochrome bj E. coli CTAB/cyclohexane/decanol Extraction [103]... 

Recombinant Cytochrome C553 E. coli periplasm AOT/isoocatne Extraction and purification [104]... 

AOT/octane Recombinant cytochrome bj Forward and back extractions were performed and the efficiency of back extraction was found to decrease at low temperature and high salt concentration [289]... 

Kalgutkar, A. S., et al. (2005). Bioactivation of the nontricyclic antidepressant nefazodone to a reactive quinone-imine species in human liver microsomes and recombinant cytochrome P450 3A4. Drug Metab. Dispos. 33 243-253. 

Karam WG, Goldstein JA, Lasker JM, et al. Human CYP2C19 is a major omeprazole 5-hydroxylase, as demonstrated with recombinant cytochrome P450 enzymes. Drug Metab Dispos 1996 24 1081-1087. 

Fig. II.9 Heme environment of (a) recombinant cytochrome c peroxidase Compound III (PDB 1DCC) [149] and (b) recombinant horseradish peroxidase C Compound in (PDB 1H57) [110]. Hydrogen bonds are depicted as filled lines and coordination bonds as dashed lines...

Giraud C, Tran A, Rey E, Vincent J, Treluyer JM, Pons G. In vitro characterization of clobazam metabolism by recombinant cytochrome P450 enzymes importance of CYP2C19. Drug Metab Dispos 2004 32(ll) 1279-86. 

Predictive models can be better produced when recombinant cytochrome data are available, an experimental technique which may increase the probability of obtaining consistent and predictive models, since one protein is involved in the metabolic reaction. The most accurate data to describe the rate and affinity of the ligand towards an enzyme are the kinetic parameters Vmax and Km. Nevertheless, the calculation of these parameters is time consuming. A less precise parameter is the determination of the compound percentage remaining in a cytochrome incubation after a certain period of time. These metabolic data are less accurate and can only be used to classify the compounds in a metabolic system as stable or unstable. This type of data was the basis for a predictive model of metabolic stability towards CYP3A4 [35]. 

Patki, K.C., von Moltke, L.L. and Greenblatt, D.J. (2003) In vitro metabolism of midazolam, triazolam, nifedipine, and testosterone by human liver microsomes and recombinant cytochromes P-450 role of CYP3A4 and CYP3A5. Drug Metabolism and Disposition The Biological Fate of Chemicals, 31, 938-944. 

Shou, M., Krausz, K. W., Gonzalez, F. J., and Gelboin, H. V. (1996) Metabolic activation of the potent carcinogen dibenzo[a,l]pyrene by human recombinant cytochrome P450, lung and liver microsomes. Carcinogenesis 17, 2429-2433. 

Taken together, these experiments indicate that human recombinant cytochromes expressed in E. coli are characterized by substrate specificities, reaction rates, turnover numbers and specific inhibition profiles in keeping with the counterpart enzymes from human liver microsomes. 

The bacterial expression of fuUy functional human recombinant cytochrome P450 oxi-doreductases has been estabhshed and proven to be a broadly versatile system. 

F.P. Guengerich, and T. Shimada (1996). Roles of cytochrome b5 in the oxidation of testosterone and nifedipine by recombinant cytochrome P450 3A4 and by human liver microsomes. Arch. Biochem. Biophys. 325, 174-182. 

Recombinant cytochrome P450 2D18 metabolism of dopamine and arachidonic acid. J. Pharmacol. Exp. Ther. 294, 1120-1130. 

Although a few studies have described medium-throughput microfluidic cell assays, the potential of this application has not yet been fully realized. Quake and coworkers [16] developed a device comprising hundreds of reaction chambers, a complex network of microchannels, and multiplexed valves. The utility of the device was illustrated by detecting a recombinant cytochrome c peroxidase (an enzyme used for chemiluminescent detection) expressed in E. coli. This device, which houses 256,725 nL chambers, demonstrates the potential for high-throughput cell assays with minimal reagent use. A similar platform was developed by Chin et al. [17] and contains 10,000 microwells (Fig. 3a). Rat neural stem... 

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