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Information on EC 3.1.3.11 - fructose-bisphosphatase and Organism(s) Sus scrofa and UniProt Accession P00636
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3.1.3.11 fructose-bisphosphataseIUBMB Comments
The animal enzyme also acts on sedoheptulose 1,7-bisphosphate.
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Word Map
- 3.1.3.11
- fbpases
-
gluconeogenesis
-
gluconeogenic
- amp
- chloroplast
- phosphoenolpyruvate
-
carboxykinase
- glycogen
- glucose-6-phosphatase
-
phosphofructokinase
- hexokinase
- 2,6-bisphosphate
- spinach
- malate
- thioredoxins
- carboxylase
- starch
- co2
- glucokinase
- phosphorylase
-
6-phosphatase
- fructose-2,6-bisphosphate
- pepck
- trx
- fructose-6-phosphate
- 3-phosphoglycerate
-
nadp-malate
- oleracea
- dihydroxyacetone
- g6pase
- ribulose
- ribulose-1,5-bisphosphate
- triose
-
calvin
- glucagon
- rubisco
-
sucrose-phosphate
-
light-activated
-
amp-dependent
-
antihyperglycemic
- phosphoribulokinase
- 6-phosphofructo-1-kinase
-
calvin-benson
-
monophosphatase
- rubp
- ribulose-5-phosphate
- biofuel production
- synthesis
- medicine
- biotechnology
- molecular biology
- drug development
- diagnostics
-
4.1.1.39
-
t-states
-
2.7.1.11
-
triose-phosphate
The taxonomic range for the selected organisms is: Sus scrofa
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
fructose-1,6-bisphosphatase, fructose 1,6-bisphosphatase, fructose bisphosphatase, fructose-bisphosphatase, fructose 1,6-diphosphatase, fbp-1, fructose diphosphatase, fru-1,6-p2ase, cytosolic fbpase, cfbp1, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
CY-F1
-
-
-
-
D-fructose 1,6-diphosphatase
-
-
-
-
D-fructose-1,6-bisphosphatase
-
-
-
-
D-fructose-1,6-bisphosphate 1-phosphohydrolase
-
-
-
-
D-fructose-1,6-bisphosphate phosphatase
-
-
-
-
FBPase
-
-
-
-
Fru-1,6-P2ase
-
-
-
-
fructose 1,6-bisphosphatase
-
-
-
-
fructose 1,6-bisphosphate 1-phosphatase
-
-
-
-
fructose 1,6-bisphosphate phosphatase
-
-
-
-
fructose 1,6-diphosphatase
-
-
-
-
fructose 1,6-diphosphate phosphatase
-
-
-
-
fructose bisphosphate phosphatase
-
-
-
-
fructose diphosphatase
-
-
-
-
fructose diphosphate phosphatase
-
-
-
-
hexose bisphosphatase
-
-
-
-
hexose diphosphatase
-
-
-
-
hexosediphosphatase
-
-
-
-
RAE-30
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
D-fructose 1,6-bisphosphate + H2O = D-fructose 6-phosphate + phosphate
loop 52-72 is an essential element in the allosteric mechanism of enzyme
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
PATHWAY SOURCE
PATHWAYS
KEGG
-
-, -, -, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
D-fructose-1,6-bisphosphate 1-phosphohydrolase
The animal enzyme also acts on sedoheptulose 1,7-bisphosphate.
CAS REGISTRY NUMBER
COMMENTARY
9001-52-9
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
binding of the first substrate molecule, in one dimer of the enzyme, produces a conformational change at the other dimer, reducing the substrate affinity and catalytic activity of its subunits
-
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
r
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
enzyme is usually regarded as a regulatory enzyme of gluconeogenesis
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
enzyme is usually regarded as a regulatory enzyme of gluconeogenesis
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
enzyme is usually regarded as a regulatory enzyme of gluconeogenesis
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-fructose 1,6-diphosphate + H2O
D-fructose 6-phosphate + phosphate
enzyme is usually regarded as a regulatory enzyme of gluconeogenesis
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
enzyme is usually regarded as a regulatory enzyme of gluconeogenesis
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
?
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
-
-
-
r
D-fructose 1,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
enzyme is usually regarded as a regulatory enzyme of gluconeogenesis
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
K+
additional information
-
monovalent cations activate by helping the Arg 276 residue "deshield" the partial negative charge on the 1-phosphoryl group of the substrate so that nucleophilic attack on the 1-phosphorus atom is facilitated
Mg2+
activates, required for activity, Ka values for wild-type and mutant enzymes, kinetics, overview
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(4-[3-(6,7-diethoxy-quinazolin-4-ylamino)-phenyl]-thiazol-2-yl)-methanol
i.e. PFE, allosteric inhibitor, residue L56 coordinates the (4-[3-(6,7-diethoxy-quinazolin-4-ylamino)-phenyl]-thiazol-2-yl)-methanol (PFE) inhibitor ligand, as does residue L73, both of which exhibit hydrophobic interactions with the ligand in the PFE-binding site. In addition, L73 and L56 are part of a network that leads from the allosteric binding site to the active site of the enzyme. This hydrophobic network, also involving residues V48 and L120 may stabilize previously described hydrogen bonding networks including residues R49, S169, and D127, shown in the network, leading to the active site where the metal binds D121, D118, and E280. The M177 and Y164 interfacial residues are positioned between the AMP-binding site and active sites
D-fructose 2,6-bisphosphate
-
the binding of fructose 1,6-bisphosphate induces the appearance of catalytic sites with lower affinity for substrate and lower catalytic activity. The inhibitor, fructose 1,6-bisphosphate, competes with the substrate for the high-affinity sites. Binding of substrate to the low-affinity sites acts as a stapler that prevents dissociation of the tetramer and hence exchange of subunits, and results in substrate inhibition
additional information
the FBPase pig kidney tetramer overlay of human and pig kidney (PDB IDs 1FTA and 1KZ8, respectively) show nearly identical orientation and conformation in the active site, AMP allosteric binding site, and inhibitor (4-[3-(6,7-diethoxy-quinazolin-4-ylamino)-phenyl]-thiazol-2-yl)-methanol allosteric binding site architecture
-
AMP
wild-type, 50% inhibition at 0.0016 mM, mutant E97A, 50% inhibition at 0.0038 mM, mutant D118A, 50% inhibition at 0.0069 mM, mutant D121A, 50% inhibition at 0.0074 mM
fructose 2,6-bisphosphate
global conformational change in porcine FBPase induced by fructose 2,6-bisphosphate in the absence of AMP
AMP
-
uptake of 1 M of NEM per mol of subunit is accompanied by a loss of sensitivity towards AMP. K+ induces a conformational change on the enzyme derivative which hinders AMP interaction with the protein
AMP
-
study of allosteric inhibition, two classes of binding sites with two distinct affinities for AMP are possible
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0042 - 0.0048
D-fructose 1,6-bisphosphate
pH 7.5, 37°C, recombinant mutant M177A
0.0048
D-fructose 1,6-bisphosphate
pH 7.5, 37°C, recombinant wild-type enzyme
0.0048
D-fructose 1,6-bisphosphate
30°C, pH 7.5, recombinant wild-type enzyme
additional information
additional information
enzyme shows a mechanism of cooperativity that arises from within a single subunit
-
additional information
additional information
-
enzyme shows a mechanism of cooperativity that arises from within a single subunit
-
additional information
additional information
-
comparison of KM of wild-type and hybrid enzymes
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
comparison of kcat of wild-type and hybrid enzymes
-
11.5 - 12.8
D-fructose 1,6-bisphosphate
pH 7.5, 37°C, recombinant mutant M177A
19.7
D-fructose 1,6-bisphosphate
30°C, pH 7.5, recombinant wild-type enzyme
20.5
D-fructose 1,6-bisphosphate
pH 7.5, 37°C, recombinant wild-type enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4271
D-fructose 1,6-bisphosphate
pH 7.5, 37°C, recombinant wild-type enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00025
D-fructose 2,6-bisphosphate
pH 7.5, 37°C, recombinant wild-type enzyme
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00062 - 0.00075
D-fructose 2,6-bisphosphate
Sus scrofa
pH 7.5, 37°C, recombinant mutant M177A
0.001
D-fructose 2,6-bisphosphate
Sus scrofa
30°C, pH 7.5, recombinant wild-type enzyme
0.0015
D-fructose 2,6-bisphosphate
Sus scrofa
pH 7.5, 37°C, recombinant wild-type enzyme
0.0017 - 0.0018
D-fructose 2,6-bisphosphate
Sus scrofa
pH 7.5, 37°C, recombinant mutant Y113A
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
enzyme FBPase is a key rate-controlling enzyme in the gluconeogenic pathway
physiological function
enzyme FBPase is a key rate-controlling enzyme in the gluconeogenic pathway. FBPase activity is regulated synergistically by the allosteric inhibitors AMP and fructose-2,6-bisphosphate (F2,6-BP). FBPase functions in the degradation of fructose-1,6-bisphosphate (FBP), which hydrolyzes to fructose-6-phosphate (F6P) and phosphate
additional information
sequence comparisons of human and porcine FBPase 1, overview. Residue L56 coordinates the (4-[3-(6,7-diethoxy-quinazolin-4-ylamino)-phenyl]-thiazol-2-yl)-methanol (PFE) inhibitor ligand, as does residue L73, both of which exhibit hydrophobic interactions with the ligand in the PFE-binding site. In addition, L73 and L56 are part of a network that leads from the allosteric binding site to the active site of the enzyme. Residue M248 is positioned near the triad of acidic residues co-ordinating manganese and is found in the active site to co-ordinate D-fructose 6-phosphate
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). F16P1_PIG
338
1
36779
Swiss-Prot
other Location (Reliability: 2)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
tetramer
additional information
additional information
structure comparisons of human and porcine kidney enzymes, overview. The FBPase pig kidney tetramer overlay of human and pig kidney (PDB IDs 1FTA and 1KZ8, respectively) show nearly identical orientation and conformation in the active site, AMP allosteric binding site, and inhibitor (4-[3-(6,7-diethoxy-quinazolin-4-ylamino)-phenyl]-thiazol-2-yl)-methanol allosteric binding site architecture
additional information
-
on molecular surface of enzyme, identification of a number of domains specific for proteins transported to the nucleus
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
construction of hybrids in which subunits have either their active or regulatory sited rendered non-functional by specific mutations
additional information
-
formation of hybrid tetramers and comparison of kinetic parameters
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drops in vapor diffusion, crystals of Zn2+/fructose 2,6-bisphosphate complex
In presence of AMP, the enzyme crystallizes in the T-state and AMP displaces loop 52-72 from its engaged conformation and abolishes metal association with sites 2 and 3. In the absence of AMP, enzyme is in the R-state and loop 52-72 associates with the active site. Three metal-binding sites are occupied by Zn2+ and two of three metal sites by Mg2+.
mutation I10D introduces an electrostatic charge that destabilizes the R and T states. Structure and molcular dynamic simulation show that the AMP/Mg2+ and AMP/Zn2+ complexes of mutant I10D are in intermediate quaternary conformations completing 12° of the subunit-pair rotation, but the complex with Zn2+ provides an engaged loop in a near-T quaternary state. The 12° subunit-pair rotation generates close contacts involving the hinges, residues 50-57 and hairpin turns, residues 58-72, of the engaged loops. Additional subunit-pair rotation toward the T state would make such contacts unfavorable
crystal structure of the enzyme complexed with AMP, the substrate analogue 2,5-anhydro-D-glucitol 1,6-diphosphosphate and Mn2+
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A51P
The mutation has little effect on the binding affinity of AMP, but increases the KI value. The KM value is unchanged.
D118A
mutant tetramer with one wild-type subunit and three mutant subunits. Kinetic parameters similar to wild-type, kcat-value is about one-fourth that of wild-type
D121A
mutant tetramer with one wild-type subunit and three mutant subunits. Kinetic parameters similar to wild-type, kcat-value is about one-fourth that of wild-type
E97A
mutant tetramer with one wild-type subunit and three mutant subunits. Kinetic parameters similar to wild-type, kcat-value is about one-fourth that of wild-type
F16W
mutant FBPases exhibits identical electrophoretic mobility as FBPase isolated from pig kidney. Mutation does not affect catalytic properties significantly, except the loss of AMP cooperativity
F219W
mutant FBPases exhibits identical electrophoretic mobility as FBPase isolated from pig kidney. Mutation does not affect catalytic properties significantly
F232W
mutant FBPases exhibits identical electrophoretic mobility as FBPase isolated from pig kidney. Mutation does not affect catalytic properties significantly
F89W
mutant FBPases exhibits identical electrophoretic mobility as FBPase isolated from pig kidney. Mutation does not affect catalytic properties significantly
G191A
decreased Km-value for fructose 1,6-diphosphate, decreased inhibition constant for fructose 1,6-diphosphate and decreased Mg2+ affinity compared to the wild type enzyme. The 50% inhibiting concentration of AMP is increased over 2000fold relative to the wild type enzyme, loss of AMP cooperativity, mechanism of AMP inhibition changes from competitive to noncompetitive
I10D
mutation introduces an electrostatic charge that destabilizes the R and T states. Structure and molcular dynamic simulation show that the AMP/Mg2+ and AMP/Zn2+ complexes of mutant I10D are in intermediate quaternary conformations completing 12° of the subunit-pair rotation, but the complex with Zn2+ provides an engaged loop in a near-T quaternary state. The 12° subunit-pair rotation generates close contacts involving the hinges, residues 50-57 and hairpin turns, residues 58-72, of the engaged loops. Additional subunit-pair rotation toward the T state would make such contacts unfavorable
I190T
decreased Km-value for fructose 1,6-diphosphate, decreased inhibition constant for fructose 1,6-diphosphate decreased Mg2+ affinity compared to the wild type enzyme. The 50% inhibiting concentration of AMP is increased over 2000fold relative to the wild type enzyme, loss of AMP cooperativity, mechanism of AMP inhibition changes from competitive to noncompetitive
K112A
K42E
decreased Km-value for fructose 1,6-diphosphate, decreased inhibition constant for fructose 1,6-diphosphate and decreased Mg2+ affinity compared to the wild type enzyme. The 50% inhibiting concentration of AMP is increased over 2000fold relative to the wild type enzyme, loss of AMP cooperativity
K42T
decreased Km-value for fructose 1,6-diphosphate, decreased inhibition constant for fructose 1,6-diphosphate decreased Mg2+ affinity compared to the wild type enzyme. The 50% inhibiting concentration of AMP is increased over 2000fold relative to the wild type enzyme, loss of AMP cooperativity, mechanism of AMP inhibition changes from competitive to noncompetitive
K50P
The mutation has little effect on the binding affinity of AMP, but increases the KI value. The KM value is unchanged, but 40fold loss if specific activity in comparison of wild-type enzyme, the Hill coefficients of Mg2+ are significantly reduced
K50P/Y57W
the KI value of AMP is increased, the mutant displays a biphasic bahavior toward AMP, the KM value is unchanged
L56A
L73A
M177A
M248D
Q32L
decreased Km-value for fructose 1,6-diphosphate and decreased inhibition constant for fructose 1,6-diphosphate compared to the wild type enzyme. 1.7fold increase in turnover number, 8fold increase in Mg2+ affinity. 8fold increase in 50% inhibiting concentration of AMP
Y113A
Y164A
D68E
-
mutation shifts the pH-optimum from pH 7.0 for the wild type enzyme to pH 8.5 for the mutant enzyme, decreased binding affinity for Mg2+ compared to wild type enzyme
D74E
-
mutation shifts the pH-optimum from pH 7.0 for the wild type enzyme to pH 8.5 for the mutant enzyme, decreased binding affinity for Mg2+ compared to wild type enzyme, no AMP cooperativity, kinetic mechanism of AMP inhibition with respect to Mg2+ is changed from competitive to noncompetitive
F219W
-
mutation introduced to allow for fluorescence measurements. At concentrations near the Km value, the substrate fructose 1,6-bispohosphate causes a 15% increase in the intrinsic fluorescence of the mutant
F232W
-
mutation introduced to allow for fluorescence measurements. The fluorescence emission of the mutant is not altered significantly by the substrate
K71A
-
mutation shifts the pH-optimum from pH 7.0 for the wild type enzyme to pH 7.5 for the mutant enzyme
K71M/K72M
-
mutation shifts the pH-optimum from pH 7.0 for the wild type enzyme to pH 7.5 for the mutant enzyme, 175fold increased inhibition constant for AMP, 2fold increased affinity for Mg2+
N64A
-
mutation shifts the pH-optimum from pH 7.0 for the wild type enzyme to pH 8.5 for the mutant enzyme, decreased binding affinity for Mg2+ compared to wild type enzyme, no AMP cooperativity, kinetic mechanism of AMP inhibition with respect to Mg2+ is changed from competitive to noncompetitive
N64Q
-
mutation shifts the pH-optimum from pH 7.0 for the wild type enzyme to pH 8.5 for the mutant enzyme, decreased binding affinity for Mg2+ compared to wild type enzyme
R49C
-
less thermostable than wild type enzyme, wild type values for turnover number and Km-value
R49D
-
less thermostable than wild type enzyme, wild type values for turnover number and Km-value, increased inhibition constant for fructose 2,6-diphosphate. Mechanism of AMP inhibition with respect to fructose 1,6-diphosphate changes from noncompetitive, wild type, to competitive. Mechanism of AMP inhibition with respect to fructose 1,6-diphosphate changes from noncompetitive, wild type, to uncompetitive. Loss of AMP cooperativity
R49L
-
less thermostable than wild type enzyme, wild type values for turnover number and Km-value, increased inhibition constant for fructose 2,6-diphosphate. Mechanism of AMP inhibition with respect to fructose 1,6-diphosphate changes from noncompetitive, wild type, to competitive. Loss of AMP cooperativity
R49M
-
enzyme is more thermostable than wild type enzyme, kinetic properties are similar to the wild type enzyme. Loss of AMP cooperativity
K112A
site-directed mutagenesis, mutation in the AMP-binding site to eliminate AMP hydrogen bonding to amino acids in the binding pocket
K112A
mutations in the AMP-binding site demonstrates an eight to nine-fold decrease in AMP inhibition
L56A
site-directed mutagenesis, interfacial mutant, that displays an about 5fold increased Ki for D-fructose 2,6-bisphosphate compared to wild-type
L56A
mutant enzyme (interfacial mutant) exhibits an increase in Ki for fructose-2,6-bisphosphate by approximately 5fold
L73A
site-directed mutagenesis, interfacial mutant, that displays an about 5fold increased Ki for D-fructose 2,6-bisphosphate compared to wild-type
L73A
mutant enzyme (interfacial mutant) exhibits an increase in Ki for fructose-2,6-bisphosphate by approximately 5fold
M177A
site-directed mutagenesis, the mutant data correlates with clinical data
M177A
mutation reveals an approximate two to three-fold decrease in inhibitory constants (Ki's) for natural inhibitors AMP and fructose-2,6-bisphosphate compared with the wild-type enzyme
M248D
site-directed mutagenesis, active site mutant, that displays an about 7fold increase in Ki for D-fructose 2,6-bisphosphate, a 4fold decrease in its apparent Km, and a 6fold increase in catalytic efficiency as compared to wild-type. The M248 residue is mutated to aspartic acid in an attempt to activate the enzyme as a means to enhance its binding affinity to the activating metals manganese and magnesium
M248D
the active site mutant enzyme displays an approximate 7fold increase in Ki for fructose-2,6-bisphosphate. The mutant enzyme displays a four-fold decrease in its apparent Michelis constant, and a six-fold increase in catalytic efficiency
Y113A
site-directed mutagenesis, mutation in the AMP-binding site to eliminate AMP hydrogen bonding to amino acids in the binding pocket
Y113A
mutations in the AMP-binding site demonstrates an eight to nine-fold decrease in AMP inhibition
Y164A
site-directed mutagenesis, the mutant data correlates with clinical data
Y164A
mutation reveals an approximate two to three-fold decrease in inhibitory constants (Ki's) for natural inhibitors AMP and fructose-2,6-bisphosphate compared with the wild-type enzyme
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
56
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His6-tagged wild-type and mutant pig kidney FBPases from Escherichia coli strain BL21(DE3) crude cell extract by dialysis and nickel affinity chromatography, cleavage of the His-tag, and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
gene FBP1, sequence comparisons of human and porcine enzymes, recombinant overexpression of C-terminally His6-tagged wild-type and mutant pig kidney FBPases in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain XL-1 Blue supercompetent cells
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
the unfolded FBPase mutants in 3.5 M guanidinium chloride are diluted to a concentration of 0.1 M guanidinium chloride the recoveries of enzymatic activity are as follows: 60.9% for F16W FBPase, 57.2% for F89W FBPase, 59.8% for F219W FBPase, and 63.6% for F232W FBPase. These results indicate that the unfolding process is not completely reversible. The reduced reversibility is similar to that observed for nonrecombinant FBPase (65.8%)
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Villeret, V.; Huang, S.; Zhang, Y.; Lipscomb, W.N.
Structural aspects of the allosteric inhibition of fructose-1,6-bisphosphatase by AMP: the binding of both the substrate analogue 2,5-anhydro-D-glucitol 1,6-bisphosphate and catalytic metal ions monitored by X-ray crystallography
Biochemistry
34
4307-4315
1995
Sus scrofa
Mendicino, J.; Abou-Issa, H.; Medicus, R.; Kratowich, N.
Fructose-1,6-bisphosphatase, phosphofructokinase, glycogen synthetase, phosphorylase, and protein kinase from swine kidney
Methods Enzymol.
42C
375-397
1975
Sus scrofa
-
Benkovic, S.J.; deMaine, M.M.
Mechanism of action of fructose 1,6-bisphosphatase
Adv. Enzymol. Relat. Areas Mol. Biol.
53
45-82
1981
Bos taurus, Canis lupus familiaris, Gallus gallus, Oryctolagus cuniculus, Ovis aries, Mus musculus, Rattus norvegicus, Phocidae, Sus scrofa
Marcus, F.; Edelstein, I.; Reardon, I.; Heinrikson, R.L.
Complete amino acid sequence of pig kidney fructose-1,6-bisphosphatase
Proc. Natl. Acad. Sci. USA
79
7161-7165
1982
Sus scrofa
Kurbanov, F.T.; Choe, J.; Honzatko, R.B.; Fromm, H.J.
Directed mutations in the poorly defined region of porcine liver fructose-1,6-bisphosphatase significantly affect catalysis and the mechanism of AMP inhibition
J. Biol. Chem.
273
17511-17516
1998
Sus scrofa
Villeret, V.; Huang, S.; Fromm, H.J.; Lipscomb, W.N.
Crystallographic evidence for the action of potassium, thallium, and lithium ions on fructose-1,6-bisphosphatase
Proc. Natl. Acad. Sci. USA
92
8916-8920
1995
Sus scrofa
Burton, V.A.; Chen, M.; Ong, W.C.; Ling, T.; Fromm, H.J.; Stayton, M.M.
High-level expression of porcine fructose-1,6-bisphosphatase in Escherichia coli: purification and characterization of the enzyme
Biochem. Biophys. Res. Commun.
192
511-517
1993
Sus scrofa
Zhang, R.; Villeret, V.; Lipscomb, W.N.; Fromm, H.J.
Kinetics and mechanism of activation and inhibition of porcine liver fructose-1,6-bisphosphatase by monovalent cations
Biochemistry
35
3038-3043
1996
Sus scrofa
Reyes, A.; Rodriguez, P.; Siebe, J.C.
The interaction of monovalent cations with fructose 1,6-bisphosphatase modified by N-ethylmaleimide and its relation with AMP inhibition
Biochem. Int.
26
347-356
1992
Sus scrofa
Shyur, L.F.; Aleshin, A.E.; Honzatko, R.B.; Fromm, H.J.
Biochemical properties of mutant and wild-type fructose-1,6-bisphosphatases are consistent with the coupling of intra- and intersubunit conformational changes in the T- and R-state transition
J. Biol. Chem.
271
33301-33307
1996
Sus scrofa (P00636), Sus scrofa
Shyur, L.F.; Poland, B.W.; Honzatko, R.B.; Fromm, H.J.
Major changes in the kinetic mechanism of AMP inhibition and AMP cooperativity attends the mutation of Arg49 in fructose-1,6-bisphosphatase
J. Biol. Chem.
272
26295-26299
1997
Sus scrofa
Choe, J.Y.; Fromm, H.J.; Honzatko, R.B.
Crystal structures of fructose 1,6-bisphosphatase: mechanism of catalysis and allosteric inhibition revealed in product complexes
Biochemistry
39
8565-8574
2000
Sus scrofa (P00636)
Kelley-Loughnane, N.; Biolsi, S.A.; Gibson, K.M.; Lu, G.; Hehir, M.J.; Phelan, P.; Kantrowitz, E.R.
Purification, kinetic studies, and homology model of Escherichia coli fructose-1,6-bisphosphatase
Biochim. Biophys. Acta
1594
6-16
2002
Escherichia coli, Sus scrofa (P00636), Sus scrofa
Zhang, F.W.; Zhao, F.K.; Xu, G.J.
Molecular cloning, expression and purification of muscle fructose-1,6-bisphosphatase from Zaocys dhumnades: the role of the N-terminal sequence in AMP activation at alkaline pH
Biol. Chem.
381
561-566
2000
Homo sapiens, Rattus norvegicus, Sus scrofa, Ptyas dhumnades
Verhees, C.H.; Akerboom, J.; Schiltz, E.; de Vos, W.M.; van der Oost, J.
Molecular and biochemical characterization of a distinct type of fructose-1,6-bisphosphatase from Pyrococcus furiosus
J. Bacteriol.
184
3401-3405
2002
Sus scrofa (P00636), Pyrococcus furiosus (Q8TZH9), Pyrococcus furiosus
Nelson, S.W.; Choe, J.Y.; Honzatko, R.B.; Fromm, H.J.
Mutations in the hinge of a dynamic loop broadly influence functional properties of fructose-1,6-bisphosphatase
J. Biol. Chem.
275
29986-29992
2000
Sus scrofa (P00636), Sus scrofa
Nelson, S.W.; Honzatko, R.B.; Fromm, H.J.
Hybrid tetramers of porcine liver fructose-1,6-bisphosphatase reveal multiple pathways of allosteric inhibition
J. Biol. Chem.
277
15539-15545
2002
Sus scrofa
Jang, H.K.; Lee, S.W.; Lee, Y.H.; Hahn, T.R.
Purification and characterization of a recombinant pea cytoplasmic fructose-1,6-bisphosphatase
Protein Expr. Purif.
28
42-48
2003
Escherichia coli, Spinacia oleracea, Sus scrofa, Pisum sativum (Q8RW99), Pisum sativum
Kelley-Loughnane, N.; Kantrowitz, E.R.
Binding of AMP to two of the four subunits of pig kidney fructose-1,6-bisphosphatase induces the allosteric transition
Proteins Struct. Funct. Genet.
44
255-261
2001
Sus scrofa
Nelson, S.W.; Honzatko, R.B.; Fromm, H.J.
Origin of cooperativity in the activation of fructose-1,6-bisphosphatase by Mg2+
J. Biol. Chem.
279
18481-18487
2004
Sus scrofa (P00636), Sus scrofa
Gizak, A.; Rakus, D.; Dzugaj, A.
Nuclear Localization of fructose 1,6-bisphosphatase in smooth muscle cells
J. Mol. Histol.
36
243-248
2005
Sus scrofa
Ludwig, H.C.; Pardo, F.N.; Asenjo, J.L.; Maureira, M.A.; Yanez, A.J.; Slebe, J.C.
Unraveling multistate unfolding of pig kidney fructose-1,6-bisphosphatase using single tryptophan mutants
FEBS J.
274
5337-5349
2007
Sus scrofa (P00636), Sus scrofa
Hines, J.K.; Chen, X.; Nix, J.C.; Fromm, H.J.; Honzatko, R.B.
Structures of mammalian and bacterial fructose-1,6-bisphosphatase reveal the basis for synergism in AMP/fructose 2,6-bisphosphate inhibition
J. Biol. Chem.
282
36121-36131
2007
Sus scrofa (P00636), Sus scrofa, Escherichia coli (P0A993), Escherichia coli
Heng, S.; Gryncel, K.R.; Kantrowitz, E.R.
A library of novel allosteric inhibitors against fructose 1,6-bisphosphatase
Bioorg. Med. Chem.
17
3916-3922
2009
Sus scrofa (P00636), Sus scrofa
Gao, Y.; Iancu, C.V.; Mukind, S.; Choe, J.Y.; Honzatko, R.B.
Mechanism of displacement of a catalytically essential loop from the active site of mammalian fructose-1,6-bisphosphatase
Biochemistry
52
5206-5216
2013
Sus scrofa (P00636)
Asenjo, J.L.; Ludwig, H.C.; Droppelmann, C.A.; Carcamo, J.G.; Concha, I.I.; Yanez, A.J.; Cardenas, M.L.; Cornish-Bowden, A.; Slebe, J.C.
Subunit interactions in pig-kidney fructose-1,6-bisphosphatase: binding of substrate induces a second class of site with lowered affinity and catalytic activity
Biochim. Biophys. Acta
1840
1798-1807
2014
Sus scrofa
Topaz, G.R.; Epiter-Smith, V.; Robolo, C.; Emad, M.; Ford, V.; Daley, J.; Byron, J.; Stieglitz, K.A.
Characterization of recombinant fructose 1,6-bisphosphatase (FBPase) gene mutations evidence of inhibition/activation of FBPase protein by gene mutation
Biosci. Rep.
39
BSR20180960
2019
Sus scrofa (P00636), Homo sapiens (P09467), Homo sapiens
Topaz, G.R.; Epiter-Smith, V.; Robolo, C.; Emad, M.; Ford, V.; Daley, J.; Byron, J.; Stieglitz, K.A.
Characterization of recombinant fructose 1,6-bisphosphatase (FBPase) gene mutations evidence of inhibition/activation of FBPase protein by gene mutation
Biosci. Rep.
39
BSR20180960
2019
Sus scrofa (P00636)
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