Noncovalent enzyme-substrate interactions in the catalytic mechanism of yeast aldose reductase.

The role of noncovalent interactions in the catalytic mechanism of aldose reductase from the yeast Candida tenuis was determined by steady-state kinetic analysis of the NADH-dependent reduction of various aldehydes, differing in hydrophobicity and the hydrogen bonding capability with the binary enzyme-NADH complex. In a series of aliphatic aldehydes, substrate hydrophobicity contributes up to 13.7 kJ/mol of binding energy. The aldehyde binding site of aldose reductase appears to be 1.4 times more hydrophobic than n-octanol and can accommodate a linear alkyl chain with at least seven methylene groups (approximately 14 A in length). Binding energy resulting from interactions at positions 3-6 of the aldehyde is distributed between increasing the catalytic constant 2.6-fold and decreasing the apparent dissociation constant 59-fold. Hydrogen bonding interactions of the enzyme nucleotide complex with the C-2(R) hydroxyl group of the aldehyde are crucial to transition state binding and contribute up to 17 kJ/mol of binding energy. A comparison of the kinetic data of yeast aldose reductase, a key enzyme in the metabolism of D-xylose, and human aldose reductase, a presumably perfect detoxification catalyst [Grimshaw, C. E. (1992) Biochemistry 31, 10139], clearly reflects these differences in physiological function.