Modulation of 6-phosphofructo-1-kinase oligomeric equilibrium by calmodulin: formation of active dimers.
Muscle 6-phospho-1-kinase (PFK) is the key regulatory enzyme of the glycolytic pathway and is a calmodulin-binding protein binding two calmodulin molecules per PFK protomer. This enzyme is characterized by a complex regulation that involves its allosteric behavior modulated by several ligands, which modulate the equilibrium between the active tetramers and the inactive dimers of the enzyme. Calmodulin is described to induce the dimerization of PFK, so inhibiting its catalytic activity. Here, we show that binding of calmodulin specifically to its higher-affinity site of PFK induce its dimerization without compromising enzyme catalytic activity forming a hitherto not described active dimmer of PFK. It is also shown that the dimerization is a Ca2+ -dependent event that responds to physiological intracellular Ca2+ concentrations and decrease the interaction of the enzyme to membrane site, which stimulate its catalytic activity. We propose that the effects of calmodulin on PFK reported here are of great physiological significance due to the response to physiological concentrations of Ca2+ and due to be in accordance to the known effects of calmodulin on cell ATP production. We also propose that calmodulin might affect the interaction of PFK to other cellular components as the cytoskeleton.
Marinho-Carvalho
Monica M
MM
Laboratório de Enzimologia e Controle do Metabolismo (LabECoM), Departamento de Fármacos, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Zancan
Patricia
P
Sola-Penna
Mauro
M
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology*; D019281:Dimerization; D004910:Erythrocyte Membrane / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D010732:Phosphofructokinase-1 / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D020836:Protein Structure, Quaternary / Q000187:drug effects; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D013997:Time Factors; D017186:Titrimetry
D002147:Calmodulin; D010732:Phosphofructokinase-1; C068652:1-phosphofructokinase; D002118:Calcium