Table 1 Kinetic data of pig kidney FBPase AMP and F2,6-bisphosphate sensitivity comparing mutants and WT*
EnzymeIC50 (µM)IC50 change (mutant/WT)
AMPF2,6-BPAMPF2,6-BP
K112A_168.5 (± 1.36)1.50 (± 0.05)9.101.00
K112A_252.3 (± 1.04)1.50 (± 0.04)7.001.00
Y113A_148.6 (± 0.97)1.80 (± 0.03)6.501.20
Y113A_255.3 (± 1.10)1.70 (± 0.06)7.401.10
Y164A2.50 (± 0.08)0.95 (± 0.08)0.330.63
M177A_13.50 (± 0.05)0.75 (± 0.06)0.470.50
M177A_23.90 (± 0.06)0.62 (± 0.07)0.520.41
L56A7.35 (± 0.09)7.50 (± 0.15)0.985.00
L73A6.95 (± 0.08)7.25 (± 0.14)0.934.83
M248D7.20 (± 0.30)10.5 (± 0.12)0.967.00
WT7.50 (± 0.30)1.50 (± 0.03)N/AN/A
  • * Concentration of FBP substrate tested for AMP and F2,6-BP IC50 varied from 10 to 50 µM. Assays were performed in triplicates and averaged. AMP inhibitor was tested from 2.5 to 80 µM and F2,6-BP inhibitor from 2.5 to 50 µM.