Temperature Derivative Spectroscopy To Monitor the Autoxidation Decay of Cytochromes P450

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• 作者：Abhinav Luthra ; Ilia G. Denisov ; Stephen G. Sligar
• 刊名：Analytical Chemistry
• 出版年：2011
• 出版时间：July 1, 2011
• 年：2011
• 卷：83
• 期：13
• 页码：5394-5399
• 全文大小：801K
• 年卷期：v.83,no.13(July 1, 2011)
• ISSN：1520-6882

文摘

Temperature derivative spectroscopy (TDS), a type of relaxation spectroscopy, is a powerful tool to study protein dynamics (Berendzen, J.; Braunstein, D. Proc. Natl. Acad. Sci. U. S. A.1990, 87, 1). We developed the version of temperature derivative spectroscopy to monitor kinetics of autoxidation of cytochromes P450 and applied it to study the properties of the oxy鈥揻errous complex of a human membrane bound P450, CYP19A1 (aromatase), and that of a bacterial soluble P450, CYP101 when bound with their most common substrates, androstenedione (AD) and camphor, respectively. TDS extends the panel of methods that can be used to monitor heme protein kinetics, providing a rapid measurement technique and enabling measurement of the autoxidation rate over a wide range of temperatures, yielding the activation energy as well as absolute reaction rate in a single experiment.