Reconstitution of human dihydroorotate dehydrogenase into lipid-detergent micelles

Human dihydroorotate dehydrogenase (DHODH) is an integral membrane protein found in mitochondria involved in the biosynthesis of pyrimidines. DHODH oxidizes dihydroorotate with the simultaneous reduction of ubiquinone Q10 and is a well-validated target for anti-inflammatory and anti-proliferative drugs. We are investigating the mechanisms by which DHODH interacts with Q10 and clinical inhibitors in the inner mitochondrial membrane (IMM) and therefore develop methods to reconstitute it into physiologically relevant membranes. We are reconstituting full-length DHODH into supported lipid bilayers mimicking the IMM from phospholipid-detergent micelles using dodecyl-D-maltoside (DDM) as the surfactant. In this SANS2D experiment, we propose to determine the structure of the lipid-DDM-DHODH micelles in solution using contrast variation.