Open AccessIdentification of lactate dehydrogenase as a mammalian pyrroloquinoline quinone (PQQ)-binding protein
Author(s) -
Mitsugu Akagawa,
Kenji Minematsu,
Takahiro Shibata,
Toshiyuki Kondo,
Takeshi Ishii,
Koji Uchida
Publication year - 2016
Publication title -
scientific reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.24
H-Index - 213
ISSN - 2045-2322
DOI - 10.1038/srep26723
Subject(s) - pyrroloquinoline quinone , nad+ kinase , biochemistry , lactate dehydrogenase , pyruvate dehydrogenase complex , chemistry , cofactor , dehydrogenase , oxidative phosphorylation , citric acid cycle , intracellular , redox , biology , metabolism , enzyme , organic chemistry
Pyrroloquinoline quinone (PQQ), a redox-active o -quinone, is an important nutrient involved in numerous physiological and biochemical processes in mammals. Despite such beneficial functions, the underlying molecular mechanisms remain to be established. In the present study, using PQQ-immobilized Sepharose beads as a probe, we examined the presence of protein(s) that are capable of binding PQQ in mouse NIH/3T3 fibroblasts and identified five cellular proteins, including l -lactate dehydrogenase (LDH) A chain, as potential mammalian PQQ-binding proteins. In vitro studies using a purified rabbit muscle LDH show that PQQ inhibits the formation of lactate from pyruvate in the presence of NADH (forward reaction), whereas it enhances the conversion of lactate to pyruvate in the presence of NAD + (reverse reaction). The molecular mechanism underlying PQQ-mediated regulation of LDH activity is attributed to the oxidation of NADH to NAD + by PQQ. Indeed, the PQQ-bound LDH oxidizes NADH, generating NAD + , and significantly catalyzes the conversion of lactate to pyruvate. Furthermore, PQQ attenuates cellular lactate release and increases intracellular ATP levels in the NIH/3T3 fibroblasts. Our results suggest that PQQ, modulating LDH activity to facilitate pyruvate formation through its redox-cycling activity, may be involved in the enhanced energy production via mitochondrial TCA cycle and oxidative phosphorylation.