Isocitrate dehydrogenase 1 (IDH1) catalyzes the reversible NADP-dependent reduction reaction of isocitrate to alpha ketoglutarate. The most common cancer-driving mutations in IDH1 affect residue R132, causing a neomorphic reaction of reducing alpha ketoglutarate to D-2- hydroxyglutarate. Previous researchers indicate that a mutation in IDH1 is linked to over 80% of lower- grade gliomas as well as secondary glioblastomas patients. Here I seek to understand how mutant IDH1 produces a new reaction that hasn’t been seen in the wild type form. After designing a mutation using site-directed mutagenesis, I expressed and purified this protein. Then, I used enzyme kinetics as well as other biophysical methods in order to test how efficiently this specific mutation produces D-2-hydroxyglutarate (D2HG). Key Term: Cancer, enzyme kinetics, IDH1, mutation
