Nanodisc reconstitution of flavin mononucleotide binding domain of cytochrome-P450-reductase enables high-resolution NMR probing

Cytochrome-P450-reductase transfers electrons to cytochrome-P450 through its flavin mononucleotide binding domain (FBD). Despite the importance of membrane-anchoring for FBD function, studies have focused on its soluble domain lacking the transmembrane-domain. Here we demonstrate that the reconstitution of FBD in nanodiscs enables high-resolution NMR measurements and renders a stable conformation. Reconstitution of the flavin mononucleotide binding domain of cyt-P450-reductase in peptide-lipid-nanodiscs provided a stable conformation that assisted in mapping transient lipid-protein interactions and line broadening through chemical shift analysis.