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Sci. Pharm. 2008, 76(2), 241-258; doi:10.3797/scipharm.0803-01 (registering DOI)

Catalytic Activity of Cytochrome P-450 using NADP+ Reduced by an Anionic Hydride Organosiloxane

1
Sección de Estudios de Posgrado e Investigación y Departamento de Bioquímica, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Distrito Federal, México
2
Departamento de Farmacología, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Distrito Federal, México
3
Departamento de Química, Facultad de Estudios Superiores Cuautitlán-Campo 1, Universidad Nacional Autónoma de México, Av. 1° de Mayo s/n, 74540, Estado de México, México
4
Departamento de Física, Escuela Superior de Física y Matemáticas, Instituto Politécnico Nacional, Av. Instituto Politécnico Nacional s/n, 07738, Distrito Federal, México
*
Author to whom correspondence should be addressed.
Received: 13 March 2008 / Accepted: 16 April 2008 / Published: 21 April 2008
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Abstract

Abstract Cytochrome P-450 (P450) catalyzes a wide variety of chemical reactions; however, its use for in vitro assays has several limitations, the most striking one is the use of the reduced nicotinamide adenine dinucleotide phosphate (NADPH) coenzyme. In this work, the P450 activity using NADP+ reduced by an anionic organosiloxane, commonly named silica hydride, was evaluated. The results showed that the reduction of NADP+with silica hydride was concentration- and time-dependent. P-450 activity was maintained when NADP+ and silica hydride were added during the reaction; however, it was lower than when commercial NADPH was employed. This is due to the ability of silica hydride to reduce P450 iron atom as corroborated by the electronic paramagnetic resonance (EPR). Furthermore, this compound possibly chelates FeII because, in its presence, the P450 affinity for aniline diminishes. However, the P450 activity was the best when NADP+ was reduced by silica hydride before the former was added to the reaction. Therefore, this system could be apt for studying biotransformation reactions.
Keywords: Cytochrome P450; Drug metabolism; Electron paramagnetic resonance; Nicotinamide adenine dinucleotide phosphate; Silica hydride Cytochrome P450; Drug metabolism; Electron paramagnetic resonance; Nicotinamide adenine dinucleotide phosphate; Silica hydride
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

MENDIETA-WEJEBE, J.E.; CORREA-BASURTO, J.; ACEVES, J.M.; RAMÍREZ-ROSALES, D.; TRUJILLO-FERRARA, J.; ZAMORANO-ULLOA, R.; ROSALES-HERNÁNDEZ, M.C. Catalytic Activity of Cytochrome P-450 using NADP+ Reduced by an Anionic Hydride Organosiloxane. Sci. Pharm. 2008, 76, 241-258.

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