biostudies-literatureKrishnarjuna BNational Institute of General Medical SciencesNIGMS NIH HHS11908-11915https://www.ebi.ac.uk/biostudies/studies/S-EPMC10851674biostudies-literatureUnknown94(34)Although polymer-based lipid nanodiscs are increasingly used in the structural studies of membrane proteins, the charge of the belt-forming polymer is a major limitation for functional reconstitution of membrane proteins possessing an opposite net charge to that of the polymer. This limitation also rules out the reconstitution of a protein-protein complex composed of oppositely charged membrane proteins. In this study, we report the first successful functional reconstitution of a membrane-bound redox complex constituting a cationic cytochrome P450 (CYP450) and an anionic cytochrome P450 reductase (CPR) in non-ionic inulin-based lipid nanodiscs. The gel-to-liquid-crystalline phase-transition temperature (<i>T</i>_m) of DMPC:DMPG (7:3 w/w) lipids in polymer nanodiscs was determinedAnalytical chemistryNon-Ionic Inulin-Based Polymer Nanodiscs Enable Functional Reconstitution of a Redox Complex Composed of Oppositely Charged CYP450 and CPR in a Lipid Bilayer Membrane.PMC10851674R35GM139572R35 GM139572R35 GM139573Krishnarjuna BAuchus RJIm SCMarte JRavula TRamamoorthy AfalseNon-Ionic Inulin-Based Polymer Nanodiscs Enable Functional Reconstitution of a Redox Complex Composed of Oppositely Charged CYP450 and CPR in a Lipid Bilayer Membrane.Although polymer-based lipid nanodiscs are increasingly used in the structural studies of membrane proteins, the charge of the belt-forming polymer is a major limitation for functional reconstitution of membrane proteins possessing an opposite net charge to that of the polymer. This limitation also rules out the reconstitution of a protein-protein complex composed of oppositely charged membrane proteins. In this study, we report the first successful functional reconstitution of a membrane-bound redox complex constituting a cationic cytochrome P450 (CYP450) and an anionic cytochrome P450 reductase (CPR) in non-ionic inulin-based lipid nanodiscs. The gel-to-liquid-crystalline phase-transition temperature (<i>T</i>_m) of DMPC:DMPG (7:3 w/w) lipids in polymer nanodiscs was determined2022-01-01T00:00:00Z2022 Aug2025-04-19T18:31:54.113Z2025-04-19T18:31:54.113ZS-EPMC108516743597741710.1021/acs.analchem.2c02489