Information on EC 4.3.1.B2 - imidazole glycerol phosphate synthase

Word Map on EC 4.3.1.B2
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
4.3.1.B2
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
imidazole glycerol phosphate synthase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-glutamine + H2O = L-glutamate + NH3
show the reaction diagram
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + L-glutamine = L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + NH3 = D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
P60595: IGP synthase glutamine amidotransferase subunit, P60664: IGP synthase cyclase subunit
P60595 and P60664
SwissProt
Manually annotated by BRENDA team
Q9X0C8: subunit HisH, Q9X0C6: subunit HisF
Q9X0C8 and Q9X0C6
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
-
the enzyme represents a junction between histidine biosynthesis and de novo purine biosynthesis
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-glutamine + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-monophosphate
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
L-glutamine + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
L-glutamine + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
L-glutamate + D-erythro-imidazole-glycerol-phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
-
-
-
-
?
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + L-glutamine
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + NH3
D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + NH4+
D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
NH4+ + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
L-glutamine + N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate + L-glutamine
L-glutamate + D-erythro-1-(imidazol-4-yl)glycerol phosphate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide
show the reaction diagram
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6-diazo-5-oxo-L-norleucine
-
-
albizziin
-
mixed type inhibition versus N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate, competitive inhibition versus L-glutamine
MgCl2
-
10 mM, activity is reduced by 22%
MnCl2
-
10 mM, activity is reduced by 58%
N1-[(5'-phosphoarabinitolyl)-formimino]-5-aminoimidazole-4-carboxamide ribonucleotide
-
competitive versus N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate, mixed type inhibition versus L-glutamine
N1-[(5'-phosphoribitolyl)-formimino]-5-aminoimidazole-4-carboxamide ribonucleotide
-
competitive versus N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0015 - 7
L-glutamine
0.00027 - 0.24
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
15 - 16
NH3
1 - 291
NH4+
additional information
additional information
-
kinetic model
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0059 - 9
L-glutamine
0.00029 - 9.1
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
8.6
NH3
Escherichia coli
-
pH 8.0, 30C, HisF subunit
4.3 - 8.8
NH4+
additional information
additional information
Escherichia coli
-
kinetic model
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00042 - 5700
L-glutamine
102
0.016 - 2800
N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
194875
0.55 - 0.57
NH3
27
0.03 - 0.581
NH4+
54
additional information
additional information
Escherichia coli
-
kinetic model
2
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.38 - 1.6
albizziin
0.00067 - 0.003
N1-[(5'-phosphoarabinitolyl)-formimino]-5-aminoimidazole-4-carboxamide ribonucleotide
0.004
N1-[(5'-phosphoribitolyl)-formimino]-5-aminoimidazole-4-carboxamide ribonucleotide
-
pH 7.0, 30C, N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
-
L-glutamine-dependent activity catalyzed by IGP synthase holoenzyme
8 - 9
-
ammonia-dependent activity catalyzed by HisF
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5
-
85% of maximal activity, L-glutamine-dependent activity catalyzed by IGP synthase holoenzyme
9.5
-
22% of maximal activity, L-glutamine-dependent activity catalyzed by IGP synthase holoenzyme
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
the production of the soluble HIS7 is highest at 25C with 1 mM isopropyl beta-D-thiogalactopyranoside and this condition is used for the large-scale protein preparations
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11600
-
1 * 11600 (HisH) + 1 * 31600 (HisF), gel filtration (Sephacryl S-100), the HisH and HisF proteins form a stable 1:1 dimeric complex that constitutes the imidazole glycerol phosphate synthase holoenzyme
21655
-
1 * 21655 (HisH) + 1 * 28457 (HisF), calculated from sequence, the HisH and HisF proteins form a stable 1:1 dimeric complex that constitutes the imidazole glycerol phosphate synthase holoenzyme
28457
-
1 * 21655 (HisH) + 1 * 28457 (HisF), calculated from sequence, the HisH and HisF proteins form a stable 1:1 dimeric complex that constitutes the imidazole glycerol phosphate synthase holoenzyme
31600
-
1 * 11600 (HisH) + 1 * 31600 (HisF), gel filtration (Sephacryl S-100), the HisH and HisF proteins form a stable 1:1 dimeric complex that constitutes the imidazole glycerol phosphate synthase holoenzyme
47500
-
gel filtration
55000
-
gel filtration
61000
-
1 * 61000, SDS-PAGE, the protein is bifunctional, representing a fusion between the N-terminal HisH domain and a C-terminal HisF domain
61082
-
1 * 61082, calculated from sequence, the protein is bifunctional, representing a fusion between the N-terminal HisH domain and a C-terminal HisF domain
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
-
-
monomer
-
1 * 61000, SDS-PAGE, the protein is bifunctional, representing a fusion between the N-terminal HisH domain and a C-terminal HisF domain; 1 * 61082, calculated from sequence, the protein is bifunctional, representing a fusion between the N-terminal HisH domain and a C-terminal HisF domain
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystallization at 20C by hanging-drop vapor diffusion from a 1:1 mixture of protein solution and reservoir solution [22%28% polyethylene glycol MME 5000, 0.1 M MES (pH 6.57.0), and 0.2 M (NH4)2SO4]. Crystals grow in 710 days to an average size of 0.12 * 0.12 * 0.08 mM. The 2.1 A crystal structure of imidazole glycerol phosphate synthase reveals extensive interaction of the glutaminase and cyclase catalytic domains. At the domain interface, the glutaminase active site points into the bottomof the (beta/alpha)8 barrel of the cyclase domain. An ammonia tunnel through the (beta/alpha)8 barrel connects the glutaminase docking site at the bottom to the cyclase active site at the top. A conserved gate of four charged residues controls access to the tunnel
-
structures of a complex of the substrate N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate and imidazole glycerol phosphate synthase covalently modified by the glutamine analogue acvicin at 2.5 A, apoenzyme at 2.4 A, and imidazole glycerol phosphate synthase inactivated by the glutamine analogue 6-diazo-5-oxo-L-norleucine at 2.5 A. Comparison of the free, acivicin-, and 6-diazo-5-oxo-L-norleucine-inactivated forms of the glutaminase active site reveals structural changes relevant to activation. The N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate-bound and -free forms of the enzyme shed light on flexible parts of the molecule with potential roles in crosstalk between active sites
-
vapor diffusion method is used to obtain crystals with a rod-like morphology of a maximum size of about 0.4 x 0.2 x 0.2 mm3. Crystal structure of the heterodimeric bienzyme ImGP synthase comprising the glutaminase subunit HisH and the cyclase subunit HisF, at 2.4 A resolution.
Q9X0C8 and Q9X0C6
crystals are obtained by the hanging-drop vapor diffusion method at 20C. Within two weeks, the crystals grow to full size (0.3 x 0.3 x 0.2 mm) in the space group P4(3)2(1)2 with unit cell parameters a = b = 82.2, c = 156.2 A. X-ray crystallographic study of the native enzyme at 2.3 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27
-
30 d, imidazole glycerol phosphate synthase holoenzyme, 50% loss of activity. The HisF and HisH proteins exhibit half-lives of less than 48 h under similar conditions
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
27C, 30 days, imidazole glycerol phosphate synthase holoenzyme, 50% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
-
P60595 and P60664
expressed in a bacterial system under the control of T7 polymerase promoter
-
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C124R
-
IGP synthases formed with Q123R mutant HisF subunit has no measurable activity with glutamine in vitro
E46G
-
IGP synthases formed with E46G mutant HisF subunit shows 2800fold reduction in the kcat/Km ratio for glutamine
Q123R
-
IGP synthases formed with Q123R mutant HisF subunit has no measurable activity with glutamine in vitro
R5H
-
IGP synthases formed with R5H mutant HisF subunit shows 1500fold reduction in the kcat/Km ratio for glutamine
D359A
-
2300fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: L-glutamine), 4fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 7.5fold decrease in kcat/Km of L-glutamine
K196A
-
0.43fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: L-glutamine), 1fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 3fold decrease in kcat/Km of L-glutamine
K196A/D359A
-
2300fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: L-glutamine), 4fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 1fold decrease in kcat/Km of L-glutamine
K258A
-
ratio of glutamine turnover to N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate turnover is 43:1 (wild-type ratio is 1:1), 2600fold decrease in kcat/KM of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate L-glutamine), 385fold decrease in kcat/Km N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 1055fold decrease in kcat/Km of L-glutamine
K258R
-
ratio of glutamine turnover to N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate turnover is 3:1 (wild-type ratio is 1:1), 20fold decrease in kcat/KM of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate L-glutamine), 125fold decrease in kcat/Km N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 45fold decrease in kcat/Km of L-glutamine
K360R
-
ratio of glutamine turnover to N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate turnover is 1:1 (identical to wild-type ratio), 1090fold decrease in kcat/KM of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate L-glutamine), 3.5fold decrease in kcat/Km N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 6.4fold decrease in kcat/Km of L-glutamine
N13A
-
130fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: L-glutamine), 3fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 350fold decrease in kcat/Km of L-glutamine
Q397A
-
7.5fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: L-glutamine), 1fold decrease in kcat/Km of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 18fold decrease in kcat/Km of L-glutamine
R239A
-
ratio of glutamine turnover to N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate turnover is 122:1 (wild-type ratio is 1:1), 860fold decrease in kcat/KM of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate L-glutamine), 5.4fold decrease in kcat/Km N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 3450fold decrease in kcat/Km of L-glutamine
R239H
-
ratio of glutamine turnover to N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate turnover is 154:1 (wild-type ratio is 1:1), 2400fold decrease in kcat/KM of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate L-glutamine), 5.2fold decrease in kcat/Km N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 1360fold decrease in kcat/Km of L-glutamine
R239K
-
ratio of glutamine turnover to N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate turnover is 40:1 (wild-type ratio is 1:1), 218fold decrease in kcat/KM of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate L-glutamine), 3.9fold decrease in kcat/Km N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 540fold decrease in kcat/Km of L-glutamine
R360A
-
ratio of glutamine turnover to N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate turnover is 3:1 (wild-type ratio is 1:1), 9.2fold decrease in kcat/KM of N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate L-glutamine), 1.4fold decrease in kcat/Km N1-[(5'-phosphoribulosyl)formimino]-5-aminoimidazole-4-carboxamido-1-beta-D-ribofuranosyl 5'-phosphate (cosubstrate: NH4+), 15fold decrease in kcat/Km of L-glutamine
T78M
-
the mutation does not impair substrate binding to the active site of HisF