Cytochrome P450 2J2 (CYP2J2) is primarily expressed in the heart and metabolizes arachidonic acid to active epoxyeicosatrienoic acid (EET). CYP2J2 was incorporated onto soluble nanodiscs, a nanoscale phospholipid bilayer, which can be self-assembled providing a native-like environment to the protein. The methods used included expression and purification of CYP2J2 and cytochrome P450 reductase (CPR), incorporating the protein onto nanodiscs and performing NADPH oxidation assays. Different concentrations of Ficoll 400 were used to crowd the protein and changes in CYP2J2 activity was measured by UV-Vis spectroscopy. Results demonstrated that increasing the Ficoll 400 concentration up to 200 mg/mL showed an increase in the activity of CYP2J2 Nanodiscs in the NADPH oxidation rate. It was suggested that the excluded volume, a property observed in macromolecular crowding, is responsible for the increase in CYP2J2 Nanodiscs activity.

Effect of macromolecular crowding on cytochrome P450 2J2 stability and activity

245th National Meeting (2013)

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