FIGURE 2.
Reductive formation of ICT and 2HG by wild-type and mutant IDH1 proteins. A, comparison of the radiochemical ([14C]αKG conversion to ICT and 2HG) assay for IDH1 in the presence and absence of a Glc-6-P dehydrogenase (G6P DH) NADPH-regenerating system. B, analysis of the products formed by IDH1 homodimers (WT/WT), IDH1WT/R132H heterodimers (WT/R132H), and IDH1(R132H) homodimers (R132H/R132H) using the radiochemical assay with the NADPH-regenerating system to simultaneously detect [14C]ICT and [14C]2HG by thin-layer chromatography as described under “Experimental Procedures.”