Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli BL21 (DE3) | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
K274L | site-directed mutagenesis, active site mutant. The residue K274 is very important for AMP analogue TNP-AMP to bind to the active site of FBPase | Homo sapiens |
K274L | equivalent activity to the wild-type enzyme. Wild-type and mutant FBPases behaved identically throughout expression and purification. When the residue K274 is mutated to L274, 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate can not bind to the active site, but can bind to the allosteric site | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate | able to bind not only to the AMP allosteric site but also to the fructose-1,6-bisphosphate active site | Homo sapiens | |
AMP | allosteric inhibition. The fluorescent AMP analogue, 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate (TNP-AMP), is used as a fluorescent probe as it is able to competitively inhibit AMP binding to the AMP allosteric site | Homo sapiens | |
fructose-2,6-bisphosphate | - |
Homo sapiens | |
additional information | the FBP active site works by stabilizing the FBPase, and the allosteric site impairs the activity of FBPase through its binding of a nonsubstrate molecule. Competitive inhibition of AMP, fructose 1,6-bisphosphate, or fructose 6-phosphate binding to FBPase with fluorescent AMP analogue, 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate (TNP-AMP)-binding FBPase | Homo sapiens | |
TNP-AMP | a fluorescent AMP analogue, 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate (TNP-AMP), is used as a fluorescent probe as it is able to competitively inhibit AMP binding to the AMP allosteric site. AMP and fructose 1,6-bisphosphate both can reduce the fluorescence from the bound TNP-AMP through competition for FBPase, suggesting that TNP-AMP binds not only to the AMP allosteric site but also to the FBP active site. The residue K274 is very important for TNP-AMP to bind to the active site of FBPase. When the residue K274 is mutated to L274, TNP-AMP cannot bind to the active site, but can bind to the allosteric site | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.018 | - |
D-fructose 1,6-bisphosphate | pH 7.4, temperature not specified in the publication, wild-type enzyme | Homo sapiens | |
0.04 | - |
D-fructose 1,6-bisphosphate | pH 7.4, temperature not specified in the publication, mutant enzyme K274L | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | activates | Homo sapiens | |
Mg2+ | Mg2+ or Mn2+ is required for activity | Homo sapiens | |
Mn2+ | activates | Homo sapiens | |
Mn2+ | Mg2+ or Mn2+ is required for activity | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose 1,6-bisphosphate + H2O | Homo sapiens | - |
D-fructose 6-phosphate + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose 1,6-bisphosphate + H2O | - |
Homo sapiens | D-fructose 6-phosphate + phosphate | - |
? | |
additional information | the FBP active site works by stabilizing the FBPase, and the allosteric site impairs the activity of FBPase through its binding of a nonsubstrate molecule. Competitive inhibition of AMP, fructose 1,6-bisphosphate, or fructose 6-phosphate binding to FBPase with fluorescent AMP analogue, 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate (TNP-AMP)-binding FBPase | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
FBPase | - |
Homo sapiens |
fructose-1,6-bisphosphatase | - |
Homo sapiens |
General Information | Comment | Organism |
---|---|---|
metabolism | regulatory enzyme in the gluconeogenesis pathway | Homo sapiens |
additional information | the FBP active site works by stabilizing the FBPase, and the allosteric site impairs the activity of FBPase through its binding of a nonsubstrate molecule. Competitive inhibition of AMP, fructose 1,6-bisphosphate, or fructose 6-phosphate binding to FBPase with fluorescent AMP analogue, 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-monophosphate (TNP-AMP)-binding FBPase | Homo sapiens |