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 M(4) and H(4) 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 M(4) 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 H(4) than M(4) isoform and that the latter interactions are weaker than with water molecules in the aqueous solution.
Institute of Applied Radiation Chemistry, Technical University of Lodz, ul. Zeromskiego 116, 90-924 Lodz, Poland.
Katarzyna
K
Swiderek
Piotr
P
Paneth
D020134:Catalytic Domain; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000737:chemistry* / Q000378:metabolism*; D019344:Lactic Acid / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D010072:Oxamic Acid / Q000737:chemistry / Q000378:metabolism*; D019289:Pyruvic Acid / Q000737:chemistry / Q000378:metabolism*
D007527:Isoenzymes; D019344:Lactic Acid; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase; D010072:Oxamic Acid