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Information on EC 2.4.2.14 - amidophosphoribosyltransferase and Organism(s) Escherichia coli and UniProt Accession P0AG16
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2.4.2.14 amidophosphoribosyltransferaseSpecify your search results
Word Map
- 2.4.2.14
- purine
-
phosphoribosylamine
-
glutamine-dependent
- glycinamide
- 6-diazo-5-oxo-l-norleucine
-
14cglycine
-
pp-ribose-p
- acivicin
- phosphoribosyltransferases
- ribotide
-
p-rib-pp
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
amidophosphoribosyltransferase, glutamine phosphoribosylpyrophosphate amidotransferase, phosphoribosyl pyrophosphate amidotransferase, amido phosphoribosyltransferase, phosphoribosylamidotransferase, 5-phosphoribosyl-1-pyrophosphate amidotransferase, gpatase, glutamine phosphoribosyl pyrophosphate amidotransferase, phosphoribosyl amidotransferase, gprat, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
5'-phosphoribosylpyrophosphate amidotransferase
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5-phosphoribosyl-1-pyrophosphate amidotransferase
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5-phosphoribosylpyrophosphate amidotransferase
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5-phosphororibosyl-1-pyrophosphate amidotransferase
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alpha-5-phosphoribosyl-1-pyrophosphate amidotransferase
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amidotransferase, phosphoribosyl pyrophosphate
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ATASE
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glutamine 5-phosphoribosylpyrophosphate amidotransferase
glutamine phosphoribosylpyrophosphate amidotransferase
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glutamine ribosylpyrophosphate 5-phosphate amidotransferase
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GPAT
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GPATase
phosphoribose pyrophosphate amidotransferase
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phosphoribosyl pyrophosphate amidotransferase
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phosphoribosyldiphosphate 5-amidotransferase
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phosphoribosylpyrophosphate glutamyl amidotransferase
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glutamine 5-phosphoribosylpyrophosphate amidotransferase
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GPATase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
5-phospho-beta-D-ribosylamine + diphosphate + L-glutamate = L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + diphosphate + L-glutamate = L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
binding of 5-phospho-alpha-D-ribose 1-diphosphate activates the enzyme by a structural change that lowers the Km for glutamine 100fold and couples glutamine hydrolysis to synthesis of 5-phospho-beta-D-ribosylamine
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5-phospho-beta-D-ribosylamine + diphosphate + L-glutamate = L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
mechanism of glutamine amide transfer
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
amino group transfer
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PATHWAY SOURCE
PATHWAYS
BRENDA
KEGG
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-, -, -
SYSTEMATIC NAME
IUBMB Comments
5-phospho-beta-D-ribosylamine:diphosphate phospho-alpha-D-ribosyltransferase (glutamate-amidating)
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CAS REGISTRY NUMBER
COMMENTARY
9031-82-7
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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
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + L-glutamate + diphosphate
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
L-glutamate + 5-phospho-beta-D-ribosylamine + diphosphate
NH3 + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + diphosphate
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + L-glutamate + diphosphate
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-
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?
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + L-glutamate + diphosphate
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-
?
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + L-glutamate + diphosphate
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-
-
?
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
L-glutamate + 5-phospho-beta-D-ribosylamine + diphosphate
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first reaction in de-novo pathway of purine biosynthesis
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?
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
L-glutamate + 5-phospho-beta-D-ribosylamine + diphosphate
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first reaction in de-novo pathway of purine biosynthesis
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?
L-glutamine + H2O
L-glutamate + NH3
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enzyme exhibits glutaminase activity in the absence of other substrates or effectors
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?
NH3 + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + diphosphate
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?
NH3 + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + diphosphate
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no activity with NH4+
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?
NH3 + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
5-phospho-beta-D-ribosylamine + diphosphate
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2.8fold higher aminotransferase activity compared to amidotransferase activity
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?
additional information
?
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GPATase-thioester-5'-phosphoribosylpyrophosphate complex model demonstrates the ammonia transfer between the active sites of GPATase in its active conformation. The ammonia channel in GPATase is a transient structural element, a pipe through which ammonia travels in the absence of an external driving potential. The ammonia tunnel in GPATase discriminates against ammonium ion
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-
?
additional information
?
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GPATase-thioester-5'-phosphoribosylpyrophosphate complex model demonstrates the ammonia transfer between the active sites of GPATase in its active conformation. The ammonia channel in GPATase is a transient structural element, a pipe through which ammonia travels in the absence of an external driving potential. The ammonia tunnel in GPATase discriminates against ammonium ion
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?
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
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
L-glutamate + 5-phospho-beta-D-ribosylamine + diphosphate
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
L-glutamate + 5-phospho-beta-D-ribosylamine + diphosphate
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first reaction in de-novo pathway of purine biosynthesis
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?
L-glutamine + 5-phospho-alpha-D-ribose 1-diphosphate + H2O
L-glutamate + 5-phospho-beta-D-ribosylamine + diphosphate
-
first reaction in de-novo pathway of purine biosynthesis
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
prototype for a metal-free amidophosphoribosyltransferase
additional information
-
non-heme iron is not present in significant amounts
additional information
-
non-heme iron is not present in significant amounts
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1-alpha-diphosphoryl-2-alpha,3-alpha-dihydroxy-4-beta-cyclopentane-methanol-5-phosphate
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competitive vs. 5-phospho-alpha-D-ribose 1-diphosphate in wild-type and P410W mutant, competition for the amidotransferase C site
2-amino-4-oxo-5-chloropentanoate
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approx. 80% inactivation of NH3-dependent activity after 30 min
5'-p-fluorosulfonylbenzoyladenosine
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inactivation follows pseudo-first order kinetic, AMP, GMP and 5-phospho-alpha-D-ribose 1-diphosphate protect
6-diazo-5-oxo-L-norleucine
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approx. 98% inactivation of amidotransferase activity after 30 min
iodoacetamide
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approx. 60% inactivation of amidotransferase activity after 30 min, very weak inactivation of NH3-dependent activity
AMP
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approx. 10 mM, complete inhibition, sigmoidal inhibition curve, competitive vs. 5-phospho-alpha-D-ribose 1-diphosphate, GMP and AMP together have a synergistic effect on inhibition
GMP
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approx. 2.5 mM, complete inhibition, highly sigmoidal inhibition curve, competitive vs. 5-phospho-alpha-D-ribose 1-diphosphate, GMP and AMP together have an synergistic effect on inhibition
additional information
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synergistic inhibition of glutaminase activity by AMP plus GMP and N3-AMP plus GMP
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additional information
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synergistic inhibition by AMP and GMP, binding of GMP to an allosteric i.e. A site and AMP to a proximal catalytic i. e. C site are necessary for synergistic inhibition
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Mg-phosphoribosyldiphosphate
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no glutamine-binding site in the absence of Mg-phosphoribosyldiphosphate
phosphoribosyl-5-phosphate
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3-4fold activation of glutaminase activity in the presence of phosphate or diphosphate
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
193
glutamine
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5-phospho-alpha-D-ribose 1-diphosphate-independent glutamine hydrolysis
additional information
additional information
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kinetic constants of Arg73 and Tyr74 mutants for basal and total glutaminase activity
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0.64
L-glutamine
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N101G mutant enzyme, 5-phospho-alpha-D-ribose 1-diphosphate-dependent glutaminase activity
1.42
L-glutamine
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N101D mutant enzyme, 5-phospho-alpha-D-ribose 1-diphosphate-dependent glutaminase activity
1.72
L-glutamine
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5-phospho-alpha-D-ribose 1-diphosphate-dependent glutamine hydrolysis
1.72
L-glutamine
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wild-type enzyme, 5-phospho-alpha-D-ribose 1-diphosphate-dependent glutaminase activity
2.43
L-glutamine
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G102A mutant enzyme, 5-phospho-alpha-D-ribose 1-diphosphate-dependent glutaminase activity
101
L-glutamine
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R73H mutant enzyme, 5-phospho-alpha-D-ribose 1-diphosphate-dependent glutaminase activity
110
L-glutamine
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R73L mutant enzyme, 5-phospho-alpha-D-ribose 1-diphosphate-dependent glutaminase activity
236
L-glutamine
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D127A mutant enzyme, 5-phospho-alpha-D-ribose 1-diphosphate-dependent glutaminase activity
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.053 - 0.058
1-alpha-diphosphoryl-2-alpha,3-alpha-dihydroxy-4-beta-cyclopentane-methanol-5-phosphate
0.053
1-alpha-diphosphoryl-2-alpha,3-alpha-dihydroxy-4-beta-cyclopentane-methanol-5-phosphate
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P410W mutant enzyme
0.058
1-alpha-diphosphoryl-2-alpha,3-alpha-dihydroxy-4-beta-cyclopentane-methanol-5-phosphate
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure in the absence of ligands at 2.0 A, and with bound AMP at 2.5 A
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crystal structure of 6-diazo-5-oxonorleucine inactivated enzyme at 2.3 A resolution
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
G331I
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50% of wild-type glutaminase activity, reduced inhibition by GMP, enhanced inhibition by AMP
K326Q
K326Q/P410W
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binding of GMP and AMP is abolished resulting in loss of inhibition
L415A
mutant has reduced transfer efficiency. The L415A GPATase mutant-thioester-5'-phosphoribosylpyrophosphate complex after 39 ps, shows the leakage of ammonia into bulk solution
N351A
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approx. 50% of wild-type glutaminase activity, completely insensitive to inhibition by GMP, partially resistent to inhibition by AMP
P410W
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reduced inhibition by AMP, strong synergistic inhibition by AMP and GMP
Y258F
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approx. 50% loss of glutaminase activity, very weak amidotransferase activity
Y329A
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normal glutaminase activity, approx. 20% amidotransferase activity, less sensitive to GMP inhibition than wild-type
Y465A
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100% of wild-type glutaminase activity, 28% of wild-type amidotransferase activity, inhibition by GMP and AMP is similar to wild-type
Y74A
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complete loss of glutaminase activity, little loss of amidotransferase activity
K326Q
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similar glutaminase and amidotransferase activity as wild-type, not inhibited by GMP
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
stable during all stages of purification provided thiols and oxygen are avoided
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
low concentrations of thiols e.g. dithiothreitol or 2-mercaptoethanol accelerate aerobic inactivation
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489747
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, heat treatment, DEAE-Sepharose, Blue Dextran-Sepharose, hydroxylapatite
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of wild-type, Y74A, Y258A, Y258F, K326Q, Y329A, G331I, N351A and Y465A mutant enzyme in Escherichia coli
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Messenger, L.J.; Zalkin, H.
Glutamine phosphoribosylpyrophosphate amidotransferase from Escherichia coli. Purification and properties
J. Biol. Chem.
254
3382-3392
1979
Escherichia coli
Holmes, E.W.
Kinetic, physical, and regulatory properties of amidophosphoribosyltransferase
Adv. Enzyme Regul.
19
215-231
1981
Bacillus subtilis, Cricetulus griseus, Columba livia, Escherichia coli, Homo sapiens, Mus musculus, Rattus norvegicus
Zalkin, H.
Structure, function, and regulation of amidophosphoribosyltransferase from prokaryotes
Adv. Enzyme Regul.
21
225-237
1983
Bacillus subtilis, Escherichia coli, Gallus gallus
Zhou, G.; Charbonneau, H.; Colman, R.F.; Zalkin, H.
Identification of sites for feedback regulation of glutamine 5-phosphoribosylpyrophosphate amidotransferase by nucleotides and relationship to residues important for catalysis
J. Biol. Chem.
268
10471-10481
1993
Escherichia coli
Zhou, G.; Smith, J.L.; Zalkin, H.
Binding of purine nucleotides to two regulatory sites results in synergistic feedback inhibition of glutamine 5-phosphoribosylpyrophosphate amidotransferase
J. Biol. Chem.
269
6784-6789
1994
Escherichia coli
Kim, J.H.; Wolle, D.; Haridas, K.; Parry, R.J.; Smith, J.L.; Zalkin, H.
A stable carbocyclic analog of 5-phosphoribosyl-1-pyrophosphate to probe the mechanism of catalysis and regulation of glutamine phosphoribosylpyrophosphate amidotransferase
J. Biol. Chem.
270
17394-17399
1995
Escherichia coli
Kim, J.H.; Krahn, J.M.; Tomchick, D.R.; Smith, J.L.; Zalkin, H.
Structure and function of the glutamine phosphoribosylpyrophosphate amidotransferase glutamine site and communication with the phosphoribosylpyrophosphate site
J. Biol. Chem.
271
15549-15557
1996
Escherichia coli
Muchmore, C.R.; Krahn, J.M.; Kim, J.H.; Zalkin, H.; Smith, J.L.
Crystal structure of glutamine phosphoribosylpyrophosphate amidotransferase from Escherichia coli
Protein Sci.
7
39-51
1998
Bacillus subtilis, Escherichia coli
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