Measurement of Electron Transfer through Cytochrome P450 Protein on Nanopillars and the Effect of Bound Substrates
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- 作者：John E. Jett ; David Lederman ; Lance A. Wollenberg ; Debin Li ; Darcy R. Flora ; Christopher D. Bostick ; Timothy S. Tracy ; Peter M. Gannett
- 刊名：The Journal of the American Chemical Society
- 出版年：2013
- 出版时间：March 13, 2013
- 年：2013
- 卷：135
- 期：10
- 页码：3834-3840
- 全文大小：326K
- 年卷期：v.135,no.10(March 13, 2013)
- ISSN：1520-5126
文摘
Electron transfer in cytochrome P450 enzymes is a fundamental process for activity. It is difficult to measure electron transfer in these enzymes because under the conditions typically used they exist in a variety of states. Using nanotechnology-based techniques, gold conducting nanopillars were constructed in an indexed array. The P450 enzyme CYP2C9 was attached to each of these nanopillars, and conductivity measurements made using conducting probe atomic force microscopy under constant force conditions. The conductivity measurements were made on CYP2C9 alone and with bound substrates, a bound substrate鈥揺ffector pair, and a bound inhibitor. Fitting of the data with the Poole鈥揊renkel model indicates a correlation between the barrier height for electron transfer and the ease of CYP2C9-mediated metabolism of the bound substrates, though the spin state of iron is not well correlated. The approach described here should have broad application to the measurement of electron transfer in P450 enzymes and other metalloenzymes.