Arg-94 is crucial to the catalysis of 3-isopropylmalate dehydrogenase from Thermus thermophilus HB8

The crucial role of Arg-94 in 3-isopropylmalate (IPM) dehydrogenase from Thermus thermophilus HB8 was elucidated by replacing the residue to lysine with site-directed mutagenesis. The kcat value of the R94K mutant enzyme for IPM was significantly reduced to 1/170 compared with that of native enzyme, whereas the Km for IPM was not much changed. It appeared that the major role of Arg-94 in exerting the enzymatic activity is not for the substrate recognition, but for the reaction catalysis, in such a way that Arg-94 facilitates stabilization of the transition-state in the decarboxylation step.