XXXV Reunião Anual da SBBqResumoID:8154


Modulation of 6-phosphofructo-1-kinase oligomeric equilibrium by calmodulin: formation of active dimmers.


Monica Mesquita Marinho-Carvalho, Patricia Zancan and Mauro Sola-Penna



Laboratório de Enzimologia e Controle do Metabolismo (LabECoM), Departamento de Fármacos, Faculdade de Farmácia, UFRJ

 


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 dimmers of the enzyme. Calmodulin is described to induce the dimmerization of PFK, so inhibiting its catalytic activity. Here we show that binding of calmodulin specifically to its higher-affinity site of PFK induce its dimmerization without compromising enzyme catalytic activity forming a hitherto not described active dimmer of PFK. It is also shown that the dimmerization is a Ca2+-dependent event that responds to physiological intracellular Ca2+ concentrations, presenting a K0.5 for dimmerization of 0.8 mM Ca2+. These effects are also dependent on calmodulin concentrations occurring within the calmodulin intracellular levels, presenting a K0.5 for dimmerization of 1.7 nM calmodulin. The calmodulin-induced dimmerization of PFK decreases in 18 % (P < 0.05, student's t-test) the interaction of the enzyme to cytoskeletal elements, more specifically the erythrocyte membrane site. 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, which we have reported to be an important instrument to control cancer cells growth.

Financial support: CNPq, FAPERJ, FAF/FECD, PRONEX, IM-INOFAR.