Substrate and cofactor specificity and selective inhibition of lactate dehydrogenase from the malarial parasite P-falciparum

Lactate dehydrogenase from the malarial parasite Plasmodium falciparum has many amino acid residues that are unique compared to any other known lactate dehydrogenase. This includes residues that define the substrate and cofactor binding sites. Nevertheless, parasite lactate dehydrogenase exhibits high specificity for pyruvic acid, even more restricted than the specificity of human lactate dehydrogenases M-4, and H-4,. Parasite lactate dehydrogenase exhibits high catalytic efficiency in the reduction of pyruvate, k(cat)/K-m,,,, = 9.0 x 10(8) min(-1) M-1. Parasite lactate dehydrogenase also exhibits similar cofactor specificity to the human isoforms in the oxidation of L-lactate with NAD(+) and with a series of NAD(+) analogs, suggesting a similar cofactor binding environment in spite of the numerous amino acid differences. Parasite lactate dehydrogenase exhibits an enhanced k(cat),, with the analog 3-acetylpyridine adenine dinucleotide (APAD(+)) whereas the human isoforms exhibit a lower k(cat),,. This differential response to APAD(+) provides the kinetic basis for the enzyme-based detection of malarial parasites. A series of inhibitors structurally related to the natural product gossypol were shown to be competitive inhibitors of the binding of NADH. Slight changes in structure produced marked changes in selectivity of inhibition of lactate dehydrogenase. 7-p-Trifluoromethylbenzyl-8-deoxyhemigossylic acid inhibited parasite lactate dehydrogenase, K-i=0.2 mu M, which was 65- and 400-fold tighter binding compared to the M-4 and H-4 isoforms of human lactate dehydrogenase. The results suggest that the cofactor site of parasite lactate dehydrogenase may be a potential target for structure-based drug design. (C) 1997 Elsevier Science B.V.