Assessment of the Use of Single Cytochrome P450 Enzymes in by M.R. Waterman, M. Hildebrand

By M.R. Waterman, M. Hildebrand

Metabolic learn and particularly that which specializes in the monooxygenase cytochrome P450 method has undergone dramatic innovative adjustments in the final decade. Availability of hugely subtle analytical thoughts has contributed to elevated molecular figuring out of biotransformation pathways. these days dressmaker medicinal drugs are computing device composable focussing on metabolic balance. The quick improvement of recombinant enzymes has, in lots of situations, changed the extensive paintings on P450 isolation from evidently taking place resources. Recombinant P450s should be expressed in cellphone strains and hence made to be had for reproducible and relentless use addressig various medical questions. This booklet describes the current place of such examine.

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The enzymes include CYPIA1, 1A2, 2C8/9110, 2C19, 2D6, 2EI, 3A4, 3A5 as weIl as epoxide hydro lase and two N-acetyltransferases (NAT1 and NAT2). We have expressed all of these enzymes in heterologous systems such as COS-1 cells. 5 Specific Probes and Tools To Study Human Cytochromes P450 The increasing availability of specific antibodies and selective substrates and inhibitors for each major human P450 enzyme provide the tools for initial studies in subcellular fractions from liver. Another recent advance in this field is the development of more efficient analytical methods such as liquid-phase ionization techniques combined with MSIMS analysis.

1 History and Structure Cytochromes P450 were identified as chromophores in the late 1950s (Garfinkle 1958; Klingenberg 1958) and later purified, characterized, and named "cytochrome P450" on the basis of a Soret absorbance band when reduced and complexed with carbon monoxide (Omura and Sato 1961, 1964a,b). The latter property is due to the presence of a noncovalently bound heme in the form of protoporphyrin IX. With the exception of bacterial P450s, aB P450s are membrane bound.

Enzyme and pigment composition of different microsomal fractions. Arch Biochem Biophys 77:493-509 Gillum JG, Israel DS, Polk RE (1993) Pharmacokinetic drug interactions with antimicrobial agents. Clin Pharmacokinet 25:450-482 Gonzalez FJ (1992a) Control of constitutively-expressed developmentally-activated rat hepatic cytochrome P450 genes. Keio J Med 41:68-75 Gonzalez FJ (1992b) Human P450s: problems and prospects. Trends Pharmacol Sei 13:346-352 Gonzalez FJ (1992c) In vitro systems for prediction of rates of drug clearance and drug-drug interactions.

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