Calmodulin upregulates skeletal muscle 6-phosphofructo-1-kinase reversing the inhibitory effects of allosteric modulators.
6-phosphofructo-1-kinase (PFK) is a calmodulin (CaM)-binding protein that plays a key role on the regulation of glycolysis. Each PFK monomer binds two CaM molecules inducing the dissociation of the active tetrameric conformation of the enzyme into dimers, thus inhibiting it. Recently, we have reported that the binding of one CaM per PFK monomer promotes the dimerization of the enzyme although maintaining its full catalytic activity. The present work aims to understand the regulatory role of these active PFK dimers induced by CaM. We show that the inhibition of PFK activity by ATP (>1 mM) is abolished in the presence of CaM. CaM decreases the affinity of PFK for its substrates, fructose-6-phophate and ATP. Moreover, CaM activates PFK in the presence of citrate and lactate, two inhibitory metabolites that induce the dimerization of PFK tetramers, as well as potentiate the stimulatory action of ADP and fructose-2,6-bisphosphate. Under all the conditions tested CaM induces the formation of PFK dimers, supporting that these CaM-bound dimers are active and less susceptible to inhibition by allosteric ligands. In the end, we suggest that CaM binding to PFK, which is stimulated by Ca(2+), represents an important way to increase the glycolytic pathway in cells.
Marinho-Carvalho
Monica M
MM
Laboratório de Enzimologia e Controle do Metabolismo (LabECoM), Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Brazil.
Costa-Mattos
Pedro Victor
PV
Spitz
Guilherme A
GA
Zancan
Patricia
P
Sola-Penna
Mauro
M
D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors; D000494:Allosteric Regulation; D002147:Calmodulin / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D010732:Phosphofructokinase-1 / Q000187:drug effects / Q000378:metabolism*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D015854:Up-Regulation
D002147:Calmodulin; D000255:Adenosine Triphosphate; D010732:Phosphofructokinase-1