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FruBPase II and ADP-PFK1 are involved in the modulation of carbon flow in the metabolism of carbohydrates in Methanosarcina acetivorans

Santiago-Martínez, M. Geovanni, Marín-Hernández, Álvaro, Gallardo-Pérez, Juan Carlos, Yoval-Sánchez, Belem, Feregrino-Mondragón, R. Daniela, Rodríguez-Zavala, José S., Pardo, J. Pablo, Moreno-Sánchez, Rafael, Jasso-Chávez, Ricardo
Archives of biochemistry and biophysics 2019 v.669 pp. 39-49
Methanosarcina acetivorans, adenosine monophosphate, adenosine triphosphate, carbohydrates, carbon, dephosphorylation, genes, messenger RNA, metabolism, metabolites, methanogens, phosphorylation, protein content, protein kinases
To enhance our understanding of the control of archaeal carbon central metabolism, a detailed analysis of the regulation mechanisms of both fructose1,6-bisphosphatase (FruBPase) and ADP-phosphofructokinase-1 (ADP-PFK1) was carried out in the methanogen Methanosarcina acetivorans. No correlations were found among the transcript levels of the MA_1152 and MA_3563 (frubpase type II and pfk1) genes, the FruBPase and ADP-PFK1 activities, and their protein contents. The kinetics of the recombinant FruBPase II and ADP-PFK1 were hyperbolic and showed simple mixed-type inhibition by AMP and ATP, respectively. Under physiological metabolite concentrations, the FruBPase II and ADP-PFK1 activities were strongly modulated by their inhibitors. To assess whether these enzymes were also regulated by a phosphorylation/dephosphorylation process, the recombinant enzymes and cytosolic-enriched fractions were incubated in the presence of commercial protein phosphatase or protein kinase. De-phosphorylation of ADP-PFK1 slightly decreased its activity (i.e. Vmax) and did not change its kinetic parameters and oligomeric state. Thus, the data indicated a predominant metabolic regulation of both FruBPase and ADP-PFK1 activities by adenine nucleotides and suggested high degrees of control on the respective pathway fluxes.