title: CYP2J2 epoxygenase membrane anchor plays an important role in facilitating electron transfer from CPR.
abstract: CYP2J2 epoxygenase is a membrane-bound cytochrome P450 primarily expressed in the heart and plays a significant role in cardiovascular diseases. The interactions of CYP2J2 with its redox partner, cytochrome P450 reductase (CPR), and with its substrates are quite complex and can have a significant effect on the kinetics of substrate metabolism. Here we show that the N-terminus of CYP2J2 plays an important role in the formation of CYP-CPR complex for subsequent electron transfer. We demonstrate that when CYP2J2-CPR are pre-incubated before the onset of reduction, the kinetics of reduction is triphasic and is of a similar order of magnitude to previously reported rates in other cytochrome P450 systems. However, when CYP2J2 and CPR form a complex during the time course of the experiment the kinetics of the fastest phase for N-terminus containing full-length CYP2J2 is 200 times faster than the kinetics of reduction of N-terminally truncated CYP2J2. Hence, we show that the N-terminus of CYP2J2 is very important to form a productive CYP-CPR complex to facilitate electron transfer.
authors.affiliation: Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: Daryl D
authors.initials: DD
authors.lastname: Meling
authors.affiliation: Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61801, United States; Medical Scholars Program, University of Illinois Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: McDougle
authors.affiliation: Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL 61801, United States; Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, IL 61801, United States; Beckman Institute for Advanced Science and Technology, University of Illinois Urbana-Champaign, Urbana, IL 61801, United States; Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, United States. Electronic address: aditidas@illinois.edu.
authors.forename: Aditi
authors.initials: A
authors.lastname: Das
mesh_terms: D000818:Animals; D000072467:Cytochrome P450 Family 2 / Q000737:chemistry* / Q000378:metabolism; D004579:Electron Transport / Q000502:physiology; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry* / Q000378:metabolism; D000072417:Protein Domains; D051381:Rats
chemical_list: D000072467:Cytochrome P450 Family 2; D009251:NADPH-Ferrihemoprotein Reductase
keywords: CYP2J2; Cytochrome P450 reductase; Electron transfer; Nanodiscs; Stopped flow