Differences and similarities in binding of pyruvate and l-lactate in the active site of M4 and H4 isoforms of human lactate dehydrogenase

We present QM/MM calculations that show differences in geometries of active sites of M4 and H4 isoforms of human LDH ligated with oxamate, pyruvate or l-lactate. As the consequence of these differences, binding isotope effects of the methyl hydrogen atoms of pyruvate and l-lactate may be used to experimentally distinguish these isoforms. Based on the FEP calculations we argue that l-lactate is a better candidate for the experimental studies. Our calculations of energies of interactions of ligands with the active site residues provide explanation for the observed experimentally sensitivity to inhibition of the M4 isoenzyme isoform and pinpoint the differences to interactions of the ligand with the histidine residue. We conclude that pyruvate interacts much stronger in the active site of H4 than M4 isoform and that the latter interactions are weaker than with water molecules in the aqueous solution.

Theoretical calculations at the DFT level indicate that hydrogen binding isotope effect of L-lactate binding to lactate dehydrogenase is the best candidate for experimental differentation of M4 and H4 isoforms of the enzyme.Figure optionsDownload high-quality image (192 K)Download as PowerPoint slideResearch highlights
► Isoforms M4 and H4 of human lactate dehydrogenase exhibit different BIEs of pyruvate and l-lactate.
► Hydrogen BIEs of l-lactate and pyruvate can be used for differentiation of the LDH isoforms.
► Affinities of pyruvate toward active sites of M4 and H4 are different.
► l-lactate is a good candidate for experimental differentiation of LDH isoforms.