Activating Compound | Comment | Organism | Structure |
---|---|---|---|
D-glucosamine | 10 mM, 40% activation of glucokinase activity | Methanococcus maripaludis |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Methanococcus maripaludis |
Crystallization (Comment) | Organism |
---|---|
molecular modeling of structure. for binding of ADP, residues M347, I431 and L441 create a hydrophobic pocket around the adenine group. R194 makes a hydrogen bond with alpha and beta phosphates, carbonyl and NH groups from V432 peptide bond make a hydrogen bond with the NH2 group of C6 and the N1 atom of adenine | Methanococcus maripaludis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ADP | strong inhibition effect in the reverse glucokinase reaction | Methanococcus maripaludis | |
AMP | strong substrate inhibition effect in the reverse glucokinase reaction | Methanococcus maripaludis | |
D-fructose | - |
Methanococcus maripaludis | |
D-fructose 6-phosphate | strong inhibition of glucokinase activity, competitive inhibion versus D-glucose as variable substrate | Methanococcus maripaludis | |
D-glucose | substrate inhibition above 200 mM. At 10 mM 20% inhibition of phosphofructokinase activity | Methanococcus maripaludis | |
EDTA | 50 mM, complete inhibition; complete loss of activity; completely abolishes activity | Methanococcus maripaludis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.065 | - |
D-fructose 6-phosphate | pH 6.5, 30°C | Methanococcus maripaludis | |
0.065 | - |
D-fructose 6-phosphate | pH 7.0, 30°C | Methanococcus maripaludis | |
0.1 | 2 | D-glucose 6-phosphate | pH 6.5, 30°C | Methanococcus maripaludis | |
0.1 | 2 | D-fructose 1,6-bisphosphate | pH 6.5, 30°C | Methanococcus maripaludis | |
0.12 | - |
D-fructose 1,6-bisphosphate | pH 7.0, 30°C | Methanococcus maripaludis | |
8 | - |
ADP | pH 6.5, 30°C | Methanococcus maripaludis | |
8 | - |
ADP | pH 7.0, 30°C | Methanococcus maripaludis | |
40 | - |
D-glucose | pH 6.5, 30°C | Methanococcus maripaludis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | 2 mM, divalent metal ion required, activation of phosphofructokinase activity with Co2+ is about 70% compared to the activation with Mg2+, activation of glucokinase activity is about 35% compared to the activation with Mg2+ | Methanococcus maripaludis | |
Co2+ | among the divalent metal cations tested, the highest activity is observed in the presence of Mg2+, although, in the presence of Co2+, Ni2+ and Mn2+, significant activity is also measured | Methanococcus maripaludis | |
Co2+ | 2 mM, about 70% of the activity with Mg2+ | Methanococcus maripaludis | |
Mg2+ | highest activity in the presence of Mg2+ | Methanococcus maripaludis | |
Mg2+ | 2 mM, divalent metal ion required, highest activity is observed in the presence of Mg2+ | Methanococcus maripaludis | |
Mg2+ | among the divalent metal cations tested, the highest activity is observed in the presence of Mg2+, although, in the presence of Co2+, Ni2+ and Mn2+, significant activity is also measured | Methanococcus maripaludis | |
Mn2+ | 2 mM, divalent metal ion required, activation of phosphofructokinase activity with Co2+ is about 25% compared to the activation with Mg2+, activation of glucokinase activity is about 20% compared to the activation with Mg2+ | Methanococcus maripaludis | |
Mn2+ | among the divalent metal cations tested, the highest activity is observed in the presence of Mg2+, although, in the presence of Co2+, Ni2+ and Mn2+, significant activity is also measured | Methanococcus maripaludis | |
Mn2+ | 2 mM, about 25% of the activity with Mg2+ | Methanococcus maripaludis | |
additional information | divalent cation required, with highest activity in the presence of Mg2+ | Methanococcus maripaludis | |
Ni2+ | 2 mM, divalent metal ion required, activation of phosphofructokinase activity with Co2+ is about 30% compared to the activation with Mg2+, activation of glucokinase activity is less than 10% compared to the activation with Mg2+ | Methanococcus maripaludis | |
Ni2+ | among the divalent metal cations tested, the highest activity is observed in the presence of Mg2+, although, in the presence of Co2+, Ni2+ and Mn2+, significant activity is also measured | Methanococcus maripaludis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + D-fructose 6-phosphate | Methanococcus maripaludis | - |
AMP + D-fructose 1,6-bisphosphate | - |
ir | |
ADP + D-glucose | Methanococcus maripaludis | the enzyme phosphorylates both D-glucose and D-fructose 6-phosphate. Binding of both substrates to the same active site. At a sugar concentration of 10 mM the acctivity with D-fructose 6-phosphate is about 75% compared to the activity with D-glucose. No activity in presence of ATP. kcat/KM for the phosphorylation of D-fructose 6-phosphate is 440fold higher than the kcat/Km for the phosphorylation of glucose. Analysis of the kcat/Km ratios shows that the glucose dephosphorylation is 2fold more effective than the phosphorylation | AMP + D-glucose 6-phosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanococcus maripaludis | Q6LXQ3 | - |
- |
Methanococcus maripaludis | Q6LXQ3 | bifunctional ADP-dependent phosphofructokinase/glucokinase, reactions of EC 2.7.1.147 and EC 2.7.1.146, respectively | - |
Purification (Comment) | Organism |
---|---|
- |
Methanococcus maripaludis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + D-fructose 6-phosphate | - |
Methanococcus maripaludis | AMP + D-fructose 1,6-bisphosphate | - |
ir | |
ADP + D-fructose 6-phosphate | the bifunctional enzyme is able to phosphorylate D-glucose and beta-D-fructose 6-phosphate. The results of molecular modeling show that both sugars are bound to the enzyme by essentially the same residues except for N203, which establishes an interaction only when the substrate is D-fructose 6-phosphate, and E79, which interacts only with glucose. The enzyme shows higher activity with glucose compared to that obtained with beta-D-fructose 6-phosphate. beta-D-Fructose 6-phosphate shows 75% of the activity measured with glucose. In the presence of ATP, no activity is detected. Phosphatase activity is 67-fold lower than the kinase activity | Methanococcus maripaludis | AMP + D-fructose 1,6-bisphosphate | - |
ir | |
ADP + D-fructose 6-phosphate | the enzyme phosphorylates both D-glucose and D-fructose 6-phosphate. Binding of both substrates to the same active site. At a sugar concentration of 10 mM the activity with D-fructose 6-phosphate is about 75% compared to the activity with D-glucose. No activity in presence of ATP. kcat/KM for the phosphorylation of D-fructose 6-phosphate is 440fold higher than the kcat/Km for the phosphorylation of glucose | Methanococcus maripaludis | AMP + D-fructose 1,6-bisphosphate | - |
ir | |
ADP + D-fructose 6-phosphate | kinetics show hyperbolic behavior | Methanococcus maripaludis | AMP + D-fructose 1,6-bisphosphate | - |
r | |
ADP + D-glucose | the enzyme phosphorylates both D-glucose and D-fructose 6-phosphate. Binding of both substrates to the same active site. At a sugar concentration of 10 mM the acctivity with D-fructose 6-phosphate is about 75% compared to the activity with D-glucose. No activity in presence of ATP. kcat/KM for the phosphorylation of D-fructose 6-phosphate is 440fold higher than the kcat/Km for the phosphorylation of glucose. Analysis of the kcat/Km ratios shows that the glucose dephosphorylation is 2fold more effective than the phosphorylation | Methanococcus maripaludis | AMP + D-glucose 6-phosphate | - |
r | |
ADP + D-glucose | the enzyme phosphorylates both D-glucose and D-fructose 6-phosphate.Binding of both substrates to the same active site. At a sugar concentration of 10 mM the acctivity with D-fructose 6-phosphate is about 75% compared to the activity with D-glucose. No activity in presence of ATP. kcat/KM for the phosphorylation of D-fructose 6-phosphate is 440fold higher than the kcat/Km for the phosphorylation of glucose. Analysis of the kcat/Km ratios shows that the glucose dephosphorylation is 2fold more effective than the phosphorylation | Methanococcus maripaludis | AMP + D-glucose 6-phosphate | - |
r | |
AMP + D-fructose 1,6-bisphosphate | - |
Methanococcus maripaludis | ADP + D-fructose 6-phosphate | - |
r | |
CDP + beta-D-fructose 6-phosphate | about 15% compared to the activity with ADP | Methanococcus maripaludis | CMP + D-fructose 1,6-bisphosphate | - |
ir | |
additional information | less than 10% activity compared to the activity with D-glucose and ADP: L-rhamnose, D-arabinose, D-lyxose, D-fucose, D-galactose, D-mannose, D-fructose, 2-deoxyglucose, D-glucosamine, D-xylose, maltose, lactose | Methanococcus maripaludis | ? | - |
? | |
additional information | bifunctional ADP-dependent phosphofructokinase/glucokinase, reactions of EC 2.7.1.147 and EC 2.7.1.146, respectively. The rate at which fructose 6-phosphate is phosphorylated is 440fold higher than the glucose phosphorylation rate | Methanococcus maripaludis | ? | - |
? | |
UDP + D-glucose | the enzyme phosphorylates both D-glucose and D-fructose 6-phosphate. Activity with UDP and D-glucose is about 20% compared to the activity with ADP and D-glucose | Methanococcus maripaludis | UMP + D-glucose 6-phosphate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
bifunctional ADP-dependent phosphofructokinase/glucokinase | - |
Methanococcus maripaludis |
MmPFK/GK | - |
Methanococcus maripaludis |
pfkC | - |
Methanococcus maripaludis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.45 | - |
D-fructose 1,6-bisphosphate | pH 7.0, 30°C | Methanococcus maripaludis | |
0.45 | - |
D-glucose 6-phosphate | pH 6.5, 30°C | Methanococcus maripaludis | |
0.45 | - |
D-fructose 1,6-bisphosphate | pH 6.5, 30°C | Methanococcus maripaludis | |
16.5 | - |
D-fructose 6-phosphate | pH 7.0, 30°C | Methanococcus maripaludis | |
16.5 | - |
D-fructose 6-phosphate | pH 6.5, 30°C | Methanococcus maripaludis | |
23 | - |
D-glucose | pH 6.5, 30°C | Methanococcus maripaludis | |
30 | - |
ADP | pH 7.0, 30°C | Methanococcus maripaludis | |
30 | - |
ADP | pH 6.5, 30°C | Methanococcus maripaludis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
- |
Methanococcus maripaludis |
6.5 | - |
phosphofructokinase activity | Methanococcus maripaludis |
7 | - |
- |
Methanococcus maripaludis |
7 | - |
glucokinase activity | Methanococcus maripaludis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 7 | pH 5.0: about 50% of maximal activity, pH 7.0: about 65% of maximal activity | Methanococcus maripaludis |
5 | 7 | pH 5.0: about 50% of maximal phosphofructokinase activity, pH 7.0: about 65% of maximal phosphofructokinase activity | Methanococcus maripaludis |
5.5 | 7.5 | pH 5.5: about 50% of maximal phosphofructokinase activity, pH 7.5: about 75% of maximal phosphofructokinase activity | Methanococcus maripaludis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.226 | - |
D-fructose 6-phosphate | pH 7.5, 30°C, glucokinase reaction | Methanococcus maripaludis | |
0.4 | - |
AMP | pH 7.5, 30°C, reverse glucokinase reaction | Methanococcus maripaludis | |
54 | - |
ADP | pH 7.0, 30°C | Methanococcus maripaludis | |
54 | - |
ADP | pH 6.5, 30°C | Methanococcus maripaludis | |
895 | - |
D-glucose | pH 7.5, 30°C | Methanococcus maripaludis |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.08 | - |
pH 7.5, 30°C, reverse glucokinase reaction | Methanococcus maripaludis | ADP | |
0.22 | - |
pH 6.5, 30°C | Methanococcus maripaludis | D-fructose 6-phosphate | |
0.22 | - |
pH 7.5, 30°C, glucokinase reaction | Methanococcus maripaludis | D-fructose 6-phosphate | |
40 | - |
pH 6.5, 30°C | Methanococcus maripaludis | D-glucose | |
40 | - |
pH 7.5, 30°C, phosphopfructokinase reaction | Methanococcus maripaludis | D-fructose |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.57 | - |
D-glucose | pH 6.5, 30°C | Methanococcus maripaludis | |
1.09 | - |
D-glucose 6-phosphate | pH 6.5, 30°C | Methanococcus maripaludis | |
3 | - |
ADP | pH 6.5, 30°C | Methanococcus maripaludis | |
3.75 | - |
ADP | pH 7.0, 30°C | Methanococcus maripaludis | |
3.75 | - |
D-fructose 1,6-bisphosphate | pH 7.0, 30°C | Methanococcus maripaludis | |
3.75 | - |
D-fructose 1,6-bisphosphate | pH 6.5, 30°C | Methanococcus maripaludis | |
253 | - |
D-fructose 6-phosphate | pH 7.0, 30°C | Methanococcus maripaludis | |
253 | - |
D-fructose 6-phosphate | pH 6.5, 30°C | Methanococcus maripaludis |