Fructose 1,6-bisphosphatase in rat liver cytosol: interactions between the effects of K+, Zn2+, Mn2+, and fructose 2,6-bisphosphate as measured in a steady-state assay.

Fructose 1,6-bisphosphatase activity was determined in rat liver cytosols using glyceraldehyde 3-phosphate as primary substrate. Fructose 1,6-bisphosphate was formed in situ and steady-state concentrations ranging from 1 to 30 microM were observed depending on the activity of fructose 1,6-bisphosphatase and the concentration of added glyceraldehyde 3-phosphate. The system was free of contaminating low-molecular-weight compounds, divalent cations, and chelators. Under these conditions, fructose 1,6-bisphosphatase was inhibited by K+ (less than or equal to 200 mM). This inhibition was due to a reduction of V and was observed in presence of low (0.4 mM) and high (5 mM) concentrations of Mg2+. In presence of 0.4 mM Mg2+, 1 microM Zn2+ inhibited fructose 1,6-bisphosphatase by 50%; the same effect was obtained with 0.3 microM Zn2+ when the system was supplemented with 100 mM KCl. On the other hand, 0.2 microM Zn2+ enhanced the inhibitory effect of K+ and decreased the concentration of K+ yielding half-maximal inhibition from 175 to 100 mM when measured at 0.4 mM Mg2+. The effect of Zn2+ on the inhibition by K+ could be abolished by Mn2+ (less than 5 microM) or by 5 mM Mg2+. One hundred millimolar K+ enhanced the inhibition of fructose 1,6-bisphosphatase by fructose 2,6-bisphosphate and changed the type of inhibition from mainly competitive to a mixed-type inhibition (increase of Km, decrease of V). Mn2+ (less than 10 microM) reduced the effect of fructose 2,6-bisphosphate, especially in the presence of K+. It is proposed that K+ and Mn2+ may play a role in the regulation of gluconeogenesis.