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Information on EC 1.7.1.13 - preQ1 synthase and Organism(s) Escherichia coli and UniProt Accession Q46920

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IUBMB Comments
The reaction occurs in the reverse direction. This enzyme catalyses one of the early steps in the synthesis of queuosine (Q-tRNA), and is followed by the action of EC 2.4.2.29, tRNA-guanosine34 transglycosylase. Queuosine is found in the wobble position of tRNAGUN in Eukarya and Bacteria and is thought to be involved in translational modulation. The enzyme is not a GTP cyclohydrolase, as was thought previously based on sequence-homology studies.
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Escherichia coli
UNIPROT: Q46920
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The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
Synonyms
nitrile reductase, 7-cyano-7-deazaguanine reductase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
queuine synthase
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
7-aminomethyl-7-carbaguanine:NADP+ oxidoreductase
The reaction occurs in the reverse direction. This enzyme catalyses one of the early steps in the synthesis of queuosine (Q-tRNA), and is followed by the action of EC 2.4.2.29, tRNA-guanosine34 transglycosylase. Queuosine is found in the wobble position of tRNAGUN in Eukarya and Bacteria [2] and is thought to be involved in translational modulation. The enzyme is not a GTP cyclohydrolase, as was thought previously based on sequence-homology studies.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one + 2 NADPH + 2 H+
2-amino-5-amino-methyl-pyrrolo[2,3-d]pyrimidin-4-one + 2 NADP+
show the reaction diagram
natural substrate
-
-
?
2-amino-5-cyanopyrrolo[2,3-d]pyrimidine + 2 NADPH + 2 H+
2-amino-5-amino-methyl-pyrrolo[2,3-d]pyrimidine + 2 NADP+
show the reaction diagram
analogue of natural substrate, poor substrate
-
-
?
5-cyanopyrrolo[2,3-d]pyrimidin-4-one + 2 NADPH + 2 H+
5-amino-methyl-pyrrolo[2,3-d]pyrimidin-4-one + 2 NADP+
show the reaction diagram
analogue of natural substrate
-
-
?
7-cyano-7-carbaguanine + 2 NADPH + 2 H+
7-aminomethyl-7-carbaguanine + 2 NADP+
show the reaction diagram
-
-
-
?
7-cyano-7-deazaguanine + 2 NADPH + 2 H+
7-aminomethyl-7-deazaguanine + 2 NADP+
show the reaction diagram
i.e. preQ0
-
-
?
7-cyano-7-deazaguanine + NADPH + H+
queuine + NADP+ + H+
show the reaction diagram
-
-
-
?
7-cyano-7-carba-2-deaminoguanine + 2 NADPH + 2 H+
7-aminomethyl-7-carba-2-deaminoguanine + 2 NADP+
show the reaction diagram
-
-
-
-
?
7-cyano-7-carbaguanine + 2 NADPH + 2 H+
7-aminomethyl-7-carbaguanine + 2 NADP+
show the reaction diagram
-
-
-
-
?
7-cyano-7-deazaguanine + 2 NADPH + 2 H+
queuine + 2 NADP+
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one + 2 NADPH + 2 H+
2-amino-5-amino-methyl-pyrrolo[2,3-d]pyrimidin-4-one + 2 NADP+
show the reaction diagram
natural substrate
-
-
?
7-cyano-7-deazaguanine + 2 NADPH + 2 H+
7-aminomethyl-7-deazaguanine + 2 NADP+
show the reaction diagram
i.e. preQ0
-
-
?
7-cyano-7-deazaguanine + 2 NADPH + 2 H+
queuine + 2 NADP+
show the reaction diagram
-
late step in biosynthesis of the modified tRNA nucleoside queuosine
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7-formyl-7-deazaguanine
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carbonyl analogue of the imine intermediate, recognized by QueF as weak ligand for binding but not as substrate for reduction or oxidation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0061
2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one
wild-type, pH 7.5, 25°C
0.176
5-cyanopyrrolo[2,3-d]pyrimidin-4-one
wild-type, pH 7.5, 25°C
0.0015
7-cyano-7-deazaguanine
or below, pH 7.0, 37°C
0.0002 - 0.006
NADPH
0.05
7-cyano-7-carba-2-deaminoguanine
-
pH 7.5, 25°C
0.001
7-cyano-7-carbaguanine
-
below 1 microM, pH 7.5, 25°C
0.0078 - 0.036
NADPH
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.11
2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one
wild-type, pH 7.5, 25°C
0.05
5-cyanopyrrolo[2,3-d]pyrimidin-4-one
wild-type, pH 7.5, 25°C
0.127
7-cyano-7-deazaguanine
pH 7.0, 37°C
0.0033 - 0.142
NADPH
0.14
7-cyano-7-carba-2-deaminoguanine
-
pH 7.5, 25°C
0.12
7-cyano-7-carbaguanine
-
pH 7.5, 25°C
0.01
NADPH
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10.8
substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant F228W, pH 7.5, 25°C
117
substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, wild-type, pH 7.5, 25°C
12.9
substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant S90A, pH 7.5, 25°C
14.3
substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant H229A, pH 7.5, 25°C
17.9
substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant H229A, pH 7.5, 25°C
3
substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant E230Q, pH 7.5, 25°C
3.8
substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant S90A, pH 7.5, 25°C
4.4
substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant E230Q, pH 7.5, 25°C
8.7
substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant F228W, pH 7.5, 25°C
90
substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, wild-type, pH 7.5, 25°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
mass spectroscopy
homodimer
2 * 71772, calculated from amino acid sequence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
construction of homology model and docking studies of natural and non-natural substrates. Residues C190 and D197 play an essential role in the catalytic mechanism
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C190A
C190S
the mutation annihilates preQ0 covalent binding and largely disrupts the nitrile-to-amine reductase activity
D197A
the variant does not take up protons in conjunction with preQ0 binding
D197H
the variant recovers proton uptake to a level almost analogous to that seen with the wildtype enzyme
D197N
complete loss of activity
E230Q
about 4% of wild-type activity for natural substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one
E89A
the variant performs primarily (more than 90%) a two-electron reduction of 7-cyano-7-deazaguanine (preQ0), releasing hydrolyzed imine (7-formyl-7-deazaguanine) as the product
E89L
the variant performs primarily (more than 90%) a two-electron reduction of 7-cyano-7-deazaguanine (preQ0), releasing hydrolyzed imine (7-formyl-7-deazaguanine) as the product
F228A
the variant performs primarily (more than 90%) a two-electron reduction of 7-cyano-7-deazaguanine (preQ0), releasing hydrolyzed imine (7-formyl-7-deazaguanine) as the product
F228W
about 8% of wild-type activity for natural substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one
H229A
I192A
the 7-cyano-7-deazaguanine (preQ0) reduction by the mutant gives 7-aminomethyl-7-deazaguanine (preQ1) and 7-formyl-7-deazaguanine at a 1:1 ratio
L191A
the 7-cyano-7-deazaguanine (preQ0) reduction by the mutant gives 7-aminomethyl-7-deazaguanine (preQ1) and 7-formyl-7-deazaguanine at a 4:1 ratio
Q89A
poor activity
Q89L
poor activity
S90A
about 10% of wild-type activity for natural substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one
C190A
-
mutation in catalytic residue, no evidence of covalent binding of substrate preQ0. Mutant displays proton uptake
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
4 h, no loss of activity
724879
9
4 h, 80% residual activity
724879
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
half-life 28.2
40
half-life 12.8
45
half-life 2.8
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70°C, 1 year, 1% residual activity
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
immobilized metal ion affinity column chromatography and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
overproduction in Escherichia coli
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Okada, N.; Noguchi, S.; Nishimura, S.; Ohgi, T.; Goto, T.; Crain, P.F.; McCloskey, J.A.
Structure determination of a nucleoside Q precursor isolated from E. coli tRNA: 7-(aminomethyl)-7-deazaguanosine
Nucleic Acids Res.
5
2289-2296
1978
Escherichia coli
Manually annotated by BRENDA team
Noguchi, S.; Yamaizumi, Z.; Ohgi, T.; Goto, T.; Nishimura, Y.; Hirota, Y.; Nishimura, S.
Isolation of Q nucleoside precursor present in tRNA of an E. coli mutant and its characterization as 7-(cyano)-7-deazaguanosine
Nucleic Acids Res.
5
4215-4223
1978
Escherichia coli
Manually annotated by BRENDA team
van Lanen, S.G.; Reader, J.S.; Swairjo, M.A.; de Crecy-Lagard, V.; Lee, B.; Iwata-Reuyl, D.
From cyclohydrolase to oxidoreductase: discovery of nitrile reductase activity in a common fold
Proc. Natl. Acad. Sci. USA
102
4264-4269
2005
Bacillus subtilis, Escherichia coli
Manually annotated by BRENDA team
Wilding, B.; Winkler, M.; Petschacher, B.; Kratzer, R.; Egger, S.; Steinkellner, G.; Lyskowski, A.; Nidetzky, B.; Gruber, K.; Klempier, N.
Targeting the substrate binding site of E. coli nitrile reductase QueF by modeling, substrate and enzyme engineering
Chemistry
19
7007-7012
2013
Escherichia coli (Q46920), Escherichia coli
Manually annotated by BRENDA team
Moeller, K.; Nguyen, G.S.; Hollmann, F.; Hanefeld, U.
Expression and characterization of the nitrile reductase queF from E. coli
Enzyme Microb. Technol.
52
129-133
2013
Escherichia coli (Q46920)
Manually annotated by BRENDA team
Gjonaj, L.; Pinkse, M.; Fernandez-Fueyo, E.; Hollmann, F.; Hanefeld, U.
Substrate and cofactor binding to nitrile reductase A mass spectrometry based study
Catal. Sci. Technol.
6
7391-7397
2016
Escherichia coli (Q46920)
-
Manually annotated by BRENDA team
Jung, J.; Czabany, T.; Wilding, B.; Klempier, N.; Nidetzky, B.
Kinetic analysis and probing with substrate analogues of the reaction pathway of the nitrile reductase QueF from Escherichia coli
J. Biol. Chem.
291
25411-25426
2016
Escherichia coli
Manually annotated by BRENDA team
Jung, J.; Braun, J.; Czabany, T.; Nidetzky, B.
Interplay of nucleophilic catalysis with proton transfer in the nitrile reductase QueF from Escherichia coli
Catal. Sci. Technol.
9
842-853
2019
Escherichia coli (Q46920)
-
Manually annotated by BRENDA team
Jung, J.; Nidetzky, B.
Evidence of a sequestered imine intermediate during reduction of nitrile to amine by the nitrile reductase QueF from Escherichia coli
J. Biol. Chem.
293
3720-3733
2018
Escherichia coli (Q46920)
Manually annotated by BRENDA team