BRENDA - Enzyme Database
show all sequences of 2.5.1.75

Escherichia coli dimethylallyl diphosphate:tRNA dimethylallyltransferase: site-directed mutagenesis of highly conserved residues

Soderberg, T.; Poulter, C.D.; Biochemistry 40, 1734-1740 (2001)

Data extracted from this reference:

Engineering
Amino acid exchange
Commentary
Organism
D153A
at least 25% of wild-type activity
Escherichia coli
D164A
increase in Km for both substrates
Escherichia coli
D42A
20fold decrease in kcat
Escherichia coli
E173A
at least 25% of wild-type activity
Escherichia coli
E229A
substantial increase in Km for RNA, dimethylallyl diphosphate becomes inhibitory at higher concentrations
Escherichia coli
F84A
at least 25% of wild-type activity
Escherichia coli
H67F
25fold increase in Km for dimethylallyl diphosphate
Escherichia coli
K23A
increase in Km for both substrates
Escherichia coli
K280A
increase in Km for both substrates
Escherichia coli
K56A
15fold increase in Km for RNA, dimethylallyl diphosphate becomes inhibitory at higher concentrations
Escherichia coli
Q253A
18fold increase in Km for RNA, dimethylallyl diphosphate becomes inhibitory at higher concentrations
Escherichia coli
Q282A
increase in both Km and kcat value
Escherichia coli
R167A
increase in Km for both substrates
Escherichia coli
R170A
increase in Km for both substrates
Escherichia coli
R213A
at least 25% of wild-type activity
Escherichia coli
R217A
increase in Km and decrease in kcat for dimethylallyl diphosphate
Escherichia coli
R281A
at least 25% of wild-type activity
Escherichia coli
S178A
at least 25% of wild-type activity
Escherichia coli
S43A
at least 25% of wild-type activity
Escherichia coli
T108A
increase in Km for both substrates
Escherichia coli
T19A
about 600fold decrease in kcat
Escherichia coli
T24A
increase in Km for both substrates
Escherichia coli
T275A
at least 25% of wild-type activity
Escherichia coli
T54A
increase in Km for both substrates
Escherichia coli
W285A
at least 25% of wild-type activity
Escherichia coli
Y111F
at least 25% of wild-type activity
Escherichia coli
Y111S
at least 25% of wild-type activity
Escherichia coli
Y47F
at least 25% of wild-type activity
Escherichia coli
Y47S
100fold decrease in kcat
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4
-
dimethylallyl diphosphate
mutant Y47S, pH 8.0
Escherichia coli
1.5
-
seventeen-base RNA oligonucleotide
mutant H67F, pH 8.0
Escherichia coli
2.2
-
dimethylallyl diphosphate
wild-type, pH 8.0
Escherichia coli
2.3
-
dimethylallyl diphosphate
mutant Q253A, pH 8.0
Escherichia coli
2.8
-
seventeen-base RNA oligonucleotide
mutant D42A, pH 8.0
Escherichia coli
2.9
-
dimethylallyl diphosphate
mutant D42A, pH 8.0
Escherichia coli
3.2
-
seventeen-base RNA oligonucleotide
wild-type, pH 8.0
Escherichia coli
3.5
-
dimethylallyl diphosphate
mutant Y47F, pH 8.0
Escherichia coli
4.4
-
seventeen-base RNA oligonucleotide
mutant Y47F, pH 8.0
Escherichia coli
7.2
-
seventeen-base RNA oligonucleotide
mutant R217A, pH 8.0
Escherichia coli
8.9
-
seventeen-base RNA oligonucleotide
mutant Y47S, pH 8.0
Escherichia coli
10
-
dimethylallyl diphosphate
mutant K56A, pH 8.0
Escherichia coli
12
-
seventeen-base RNA oligonucleotide
mutant T54A, pH 8.0
Escherichia coli
13
-
dimethylallyl diphosphate
mutant T54A, pH 8.0
Escherichia coli
16
-
seventeen-base RNA oligonucleotide
mutant T108A, pH 8.0
Escherichia coli
22
-
dimethylallyl diphosphate
mutant T108A, pH 8.0
Escherichia coli
27
-
dimethylallyl diphosphate
mutant d164A, pH 8.0
Escherichia coli
27
-
seventeen-base RNA oligonucleotide
mutant d164A, pH 8.0
Escherichia coli
45
-
seventeen-base RNA oligonucleotide
mutant Q282A, pH 8.0
Escherichia coli
50
-
dimethylallyl diphosphate
mutant H67F, pH 8.0
Escherichia coli
50
-
seventeen-base RNA oligonucleotide
mutant K56A, pH 8.0
Escherichia coli
56
-
seventeen-base RNA oligonucleotide
mutant Q253A, pH 8.0
Escherichia coli
60
-
dimethylallyl diphosphate
mutant Q282A, pH 8.0
Escherichia coli
75
-
seventeen-base RNA oligonucleotide
mutant R167A, pH 8.0
Escherichia coli
114
-
dimethylallyl diphosphate
mutant R217A, pH 8.0
Escherichia coli
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
-
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
dimethylallyl diphosphate + seventeen-base RNA oligonucleotide
sequence 5'GCGGACUCAAAAUCCGC3', RNA oligonucleotide based on the unmodified stem-loop region of tRNAPhe
696197
Escherichia coli
diphosphate + seventeen-base RNA oligonucleotide containing 6-dimethylallyladenosine
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.002
-
seventeen-base RNA oligonucleotide
mutant Y47S, pH 8.0
Escherichia coli
0.007
-
seventeen-base RNA oligonucleotide
mutant R217A, pH 8.0
Escherichia coli
0.009
-
seventeen-base RNA oligonucleotide
mutant D42A, pH 8.0
Escherichia coli
0.07
-
seventeen-base RNA oligonucleotide
mutant T108A, pH 8.0; mutant T54A, pH 8.0
Escherichia coli
0.15
-
seventeen-base RNA oligonucleotide
mutant D164A, pH 8.0
Escherichia coli
0.18
-
seventeen-base RNA oligonucleotide
mutant H67F, pH 8.0
Escherichia coli
0.19
-
seventeen-base RNA oligonucleotide
wild-type, pH 8.0
Escherichia coli
0.23
-
seventeen-base RNA oligonucleotide
mutant Y47F, pH 8.0
Escherichia coli
0.3
-
seventeen-base RNA oligonucleotide
mutnat Q282A, pH 8.0
Escherichia coli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D153A
at least 25% of wild-type activity
Escherichia coli
D164A
increase in Km for both substrates
Escherichia coli
D42A
20fold decrease in kcat
Escherichia coli
E173A
at least 25% of wild-type activity
Escherichia coli
E229A
substantial increase in Km for RNA, dimethylallyl diphosphate becomes inhibitory at higher concentrations
Escherichia coli
F84A
at least 25% of wild-type activity
Escherichia coli
H67F
25fold increase in Km for dimethylallyl diphosphate
Escherichia coli
K23A
increase in Km for both substrates
Escherichia coli
K280A
increase in Km for both substrates
Escherichia coli
K56A
15fold increase in Km for RNA, dimethylallyl diphosphate becomes inhibitory at higher concentrations
Escherichia coli
Q253A
18fold increase in Km for RNA, dimethylallyl diphosphate becomes inhibitory at higher concentrations
Escherichia coli
Q282A
increase in both Km and kcat value
Escherichia coli
R167A
increase in Km for both substrates
Escherichia coli
R170A
increase in Km for both substrates
Escherichia coli
R213A
at least 25% of wild-type activity
Escherichia coli
R217A
increase in Km and decrease in kcat for dimethylallyl diphosphate
Escherichia coli
R281A
at least 25% of wild-type activity
Escherichia coli
S178A
at least 25% of wild-type activity
Escherichia coli
S43A
at least 25% of wild-type activity
Escherichia coli
T108A
increase in Km for both substrates
Escherichia coli
T19A
about 600fold decrease in kcat
Escherichia coli
T24A
increase in Km for both substrates
Escherichia coli
T275A
at least 25% of wild-type activity
Escherichia coli
T54A
increase in Km for both substrates
Escherichia coli
W285A
at least 25% of wild-type activity
Escherichia coli
Y111F
at least 25% of wild-type activity
Escherichia coli
Y111S
at least 25% of wild-type activity
Escherichia coli
Y47F
at least 25% of wild-type activity
Escherichia coli
Y47S
100fold decrease in kcat
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4
-
dimethylallyl diphosphate
mutant Y47S, pH 8.0
Escherichia coli
1.5
-
seventeen-base RNA oligonucleotide
mutant H67F, pH 8.0
Escherichia coli
2.2
-
dimethylallyl diphosphate
wild-type, pH 8.0
Escherichia coli
2.3
-
dimethylallyl diphosphate
mutant Q253A, pH 8.0
Escherichia coli
2.8
-
seventeen-base RNA oligonucleotide
mutant D42A, pH 8.0
Escherichia coli
2.9
-
dimethylallyl diphosphate
mutant D42A, pH 8.0
Escherichia coli
3.2
-
seventeen-base RNA oligonucleotide
wild-type, pH 8.0
Escherichia coli
3.5
-
dimethylallyl diphosphate
mutant Y47F, pH 8.0
Escherichia coli
4.4
-
seventeen-base RNA oligonucleotide
mutant Y47F, pH 8.0
Escherichia coli
7.2
-
seventeen-base RNA oligonucleotide
mutant R217A, pH 8.0
Escherichia coli
8.9
-
seventeen-base RNA oligonucleotide
mutant Y47S, pH 8.0
Escherichia coli
10
-
dimethylallyl diphosphate
mutant K56A, pH 8.0
Escherichia coli
12
-
seventeen-base RNA oligonucleotide
mutant T54A, pH 8.0
Escherichia coli
13
-
dimethylallyl diphosphate
mutant T54A, pH 8.0
Escherichia coli
16
-
seventeen-base RNA oligonucleotide
mutant T108A, pH 8.0
Escherichia coli
22
-
dimethylallyl diphosphate
mutant T108A, pH 8.0
Escherichia coli
27
-
dimethylallyl diphosphate
mutant d164A, pH 8.0
Escherichia coli
27
-
seventeen-base RNA oligonucleotide
mutant d164A, pH 8.0
Escherichia coli
45
-
seventeen-base RNA oligonucleotide
mutant Q282A, pH 8.0
Escherichia coli
50
-
dimethylallyl diphosphate
mutant H67F, pH 8.0
Escherichia coli
50
-
seventeen-base RNA oligonucleotide
mutant K56A, pH 8.0
Escherichia coli
56
-
seventeen-base RNA oligonucleotide
mutant Q253A, pH 8.0
Escherichia coli
60
-
dimethylallyl diphosphate
mutant Q282A, pH 8.0
Escherichia coli
75
-
seventeen-base RNA oligonucleotide
mutant R167A, pH 8.0
Escherichia coli
114
-
dimethylallyl diphosphate
mutant R217A, pH 8.0
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
dimethylallyl diphosphate + seventeen-base RNA oligonucleotide
sequence 5'GCGGACUCAAAAUCCGC3', RNA oligonucleotide based on the unmodified stem-loop region of tRNAPhe
696197
Escherichia coli
diphosphate + seventeen-base RNA oligonucleotide containing 6-dimethylallyladenosine
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.002
-
seventeen-base RNA oligonucleotide
mutant Y47S, pH 8.0
Escherichia coli
0.007
-
seventeen-base RNA oligonucleotide
mutant R217A, pH 8.0
Escherichia coli
0.009
-
seventeen-base RNA oligonucleotide
mutant D42A, pH 8.0
Escherichia coli
0.07
-
seventeen-base RNA oligonucleotide
mutant T108A, pH 8.0; mutant T54A, pH 8.0
Escherichia coli
0.15
-
seventeen-base RNA oligonucleotide
mutant D164A, pH 8.0
Escherichia coli
0.18
-
seventeen-base RNA oligonucleotide
mutant H67F, pH 8.0
Escherichia coli
0.19
-
seventeen-base RNA oligonucleotide
wild-type, pH 8.0
Escherichia coli
0.23
-
seventeen-base RNA oligonucleotide
mutant Y47F, pH 8.0
Escherichia coli
0.3
-
seventeen-base RNA oligonucleotide
mutnat Q282A, pH 8.0
Escherichia coli
Other publictions for EC 2.5.1.75
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
737708
Subedi
Steady-state kinetics and spec ...
Salmonella enterica, Salmonella enterica ATCC 700720
Biochemistry
54
363-376
2015
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1
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6
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1
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1
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1
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6
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1
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1
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3
3
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737635
Fradejas
Mammalian Trit1 is a tRNA([Ser ...
Mus musculus
Biochem. J.
450
427-432
2013
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1
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3
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1
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2
2
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723760
Lamichhane
Plasticity and diversity of tR ...
Homo sapiens, Saccharomyces cerevisiae, Schizosaccharomyces pombe
RNA
17
1846-1857
2011
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6
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9
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11
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6
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11
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705984
Chu
Crystal structure and substrat ...
Humulus lupulus
Nucleic Acids Res.
38
1738-1748
2010
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1
1
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1
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5
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1
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706187
Ma
Expression of isopentenyl tran ...
Agrobacterium tumefaciens
Plant Cell Rep.
28
1759-1765
2009
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1
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700955
Zhou
Crystallographic snapshots of ...
Saccharomyces cerevisiae
Proc. Natl. Acad. Sci. USA
105
16142-16147
2008
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1
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2
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1
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706146
Verdonk
Flower-specific expression of ...
Agrobacterium tumefaciens
Plant Biotechnol. J.
6
694-701
2008
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2
1
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706864
Peng
Molecular cloning of a putativ ...
Malus hupehensis
Tree Physiol.
28
899-904
2008
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3
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1
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1
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676732
Yevdakova
Characterisation of a prokaryo ...
Physcomitrella patens
Planta
226
683-695
2007
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1
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1
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1
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6
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1
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686911
Golovko
-
Expression of a human tRNA iso ...
Homo sapiens
Funct. Plant Biol.
34
654-661
2007
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1
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1
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1
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1
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689971
Kaminska
Structural bioinformatics anal ...
Salmonella enterica subsp. enterica serovar Typhimurium
Proteins Struct. Funct. Bioinform.
70
1-18
2007
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2
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676830
Miyawaki
Roles of Arabidopsis ATP/ADP i ...
Arabidopsis thaliana
Proc. Natl. Acad. Sci. USA
103
16598-16603
2006
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660041
Spinola
Identification and functional ...
Homo sapiens
Oncogene
24
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660174
Miyawaki
Expression of cytokinin biosyn ...
Arabidopsis thaliana
Plant J.
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2004
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489963
Golovko
Identification of a tRNA isope ...
Arabidopsis thaliana
Plant Mol. Biol.
49
161-169
2002
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6
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489966
Lemieux
Regulation of physiological ra ...
Caenorhabditis elegans
Genetics
159
147-157
2001
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696197
Soderberg
Escherichia coli dimethylallyl ...
Escherichia coli
Biochemistry
40
1734-1740
2001
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29
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25
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9
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29
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9
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489967
Golovko
Cloning of a human tRNA isopen ...
Homo sapiens
Gene
258
85-93
2000
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696195
Soderberg
Escherichia coli dimethylallyl ...
Escherichia coli
Biochemistry
39
6546-6553
2000
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9
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489965
Leung
Regulation of substrate recogn ...
Escherichia coli
J. Biol. Chem.
272
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1997
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2
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4
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489968
Motorin
Transfer RNA recognition by th ...
Escherichia coli
RNA
3
721-733
1997
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696190
Moore
Escherichia coli dimethylallyl ...
Escherichia coli
Biochemistry
36
604-614
1997
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2
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2
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2
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489964
Gray
Transfer RNA is the source of ...
Agrobacterium tumefaciens
Plant Physiol.
110
431-438
1996
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489962
Holtz
tRNA isopentenyltransferase fr ...
Zea mays
Hoppe-Seyler's Z. Physiol. Chem.
359
89-101
1978
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2
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5
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5
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5
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489961
Holtz
tRNA isopentenyltransferase fr ...
Lactobacillus acidophilus
Hoppe-Seyler's Z. Physiol. Chem.
356
1459-1464
1975
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2
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489959
Rosenbaum
DELTA 2 -isopentenylpyrophosph ...
Escherichia coli
J. Biol. Chem.
247
5675-5680
1972
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3
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489960
Bartz
N6-(2-isopentenyl) adenosine: ...
Escherichia coli
Biochimie
54
31-39
1972
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-
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1
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489958
Kline
N6-(delta-2-isopentenyl) adeno ...
Saccharomyces cerevisiae
Biochemistry
8
4361-4371
1969
-
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4
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2
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5
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1
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4
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