The crystal structures of mutated 3-isopropylmalate dehydrogenase from Thermus thermophilus HB8 and their relationship to the thermostability of the enzyme.

The structures of two mutant forms (G240A and L246E/V249M) of 3-isopropylmalate dehydrogenase from Thermus thermophilus HB8 were studied by X-ray crystallography. In the case of G240A, the replacement of glycine by alanine at residue 240 was expected to decrease the thermostability as a result of abnormal contacts between the methyl group of alanine and the peptide chain. However, the normal van der Waals' contacts were achieved owing to a shift in a bundle of beta-strands that yielded a vacant space for the alanine residue. The extended hydrogen bonds within the beta-sheet are the major reason for the decreased thermostability of G240A. The mutations in L246E/V249M are located in an alpha-helix region which is involved in subunit-subunit contact via hydrophobic interaction. Loosening of the subunit-subunit contact owing to ionic repulsion was the major cause of the lower heat stability of L246E/V249M.