Information on EC 6.1.1.5 - isoleucine-tRNA ligase

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

EC NUMBER
COMMENTARY hide
6.1.1.5
-
RECOMMENDED NAME
GeneOntology No.
isoleucine-tRNA ligase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + L-isoleucine + tRNAIle = AMP + diphosphate + L-isoleucyl-tRNAIle
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aminacylation
Aminoacylation
deacylation
esterification
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
-
isoleucine metabolism
-
-
tRNA charging
-
-
SYSTEMATIC NAME
IUBMB Comments
L-isoleucine:tRNAIle ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY hide
9030-96-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain 168
UniProt
Manually annotated by BRENDA team
strain 168
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain EM20031
-
-
Manually annotated by BRENDA team
K12
-
-
Manually annotated by BRENDA team
MRE600
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
the Methanopyrus kandleri sequence is compared to the sequences of the Methanopyrus isolates GC34 and GC37 from the Pacific ocean and KOL6, TAG1, TAG11, and SNP6 from the Atlantic ocean
-
-
Manually annotated by BRENDA team
Methanothermobacter thermautotrophicum
Methanothermobacter thermautotrophicum Marburg
strain Marburg
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
two isozymes, an apicoplast isozyme encoded by gene PFL1210w, and a cytoplasmic isozyme encoded by gene PF13_0179
-
-
Manually annotated by BRENDA team
gene ileS
-
-
Manually annotated by BRENDA team
strain WCUH29
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + L-isoleucine + tRNAIle
AMP + diphosphate + L-isoleucyl-tRNAIle
show the reaction diagram
ATP + L-valine + tRNAIle
AMP + diphosphate + L-valyl-tRNAIle
show the reaction diagram
-
CP1 domain of the enzyme deacylates or edits the mischarged Val-tRNAIle
r
Formycin 5'-triphosphate + isoleucine + tRNAIle
?
show the reaction diagram
-
-
-
-
-
Ile-tRNAIle + 3-mercaptopropionate
S-Isoleucyl-3-mercaptopropionate + tRNAIle
show the reaction diagram
-
-
-
-
Ile-tRNAIle + cysteamine
tRNAIle + isoleucylcysteamine
show the reaction diagram
-
-
-
-
Ile-tRNAIle + cysteine
tRNAIle + isoleucylcysteine
show the reaction diagram
-
D- and L-isomer of Lys
D-isoleucylcysteine and L-isoleucylcysteine
-
Ile-tRNAIle + DTT
Thioester of Ile and DTT + tRNAIle
show the reaction diagram
-
-
-
-
Ile-tRNAIle + L-cysteine methyl ester
tRNAIle + isoleucyl-L-cysteine methyl ester
show the reaction diagram
-
-
-
-
Ile-tRNAIle + N-acetylcysteine
S-isoleucyl-N-acetylcysteine + tRNAIle
show the reaction diagram
-
-
-
-
Tubercidin 5'-triphosphate + isoleucine + tRNAIle
?
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
ATP + L-isoleucine + tRNAIle
AMP + diphosphate + L-isoleucyl-tRNAIle
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ce3+
-
activates
Cobalt
-
cobalt-substituted enzyme is active
Dy3+
-
activates
Eu3+
-
activates
Gd3+
-
activates
La3+
-
activates
Nd3+
-
activates
Pr3+
-
activates
Sm3+
-
activates
Tb3+
-
activates
Yb3+
-
activates
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2E,4E)-5-[(2S,3R,6S,8R,9S)-3-butyl-3-[(3-carboxypropanoyl)oxy]-8-(2-hydroxyethyl)-9-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-3-methylpenta-2,4-dienoic acid
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): not determined
(2E,4E)-5-[(2S,3R,6S,8R,9S)-3-butyl-3-[(3-carboxypropanoyl)oxy]-8-[(2E)-4-hydroxy-3-methylbut-2-en-1-yl]-9-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-3-methylpenta-2,4-dienoic acid
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): not determined
(2E,4E)-5-[(2S,3R,6S,8R,9S)-3-butyl-3-[(3-carboxypropanoyl)oxy]-8-[(2E)-4-[(3-carboxypropanoyl)oxy]-3-methylbut-2-en-1-yl]-9-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-3-methylpenta-2,4-dienoic acid
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): above 15
(2E,4E)-5-[(2S,3R,6S,8R,9S)-3-butyl-3-[(3-carboxypropanoyl)oxy]-8-[(2E,4E)-6-hydroxy-3-methylhexa-2,4-dien-1-yl]-9-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-3-methylpenta-2,4-dienoic acid
-
IC50 (ng/ml): 560.3. Cell death inducibility of osteoclasts (microgram/ml): not determined
(2E,4E)-5-[(2S,3R,6S,8R,9S)-3-butyl-3-[(3-carboxypropanoyl)oxy]-8-[(2E,4E,6S,7S)-6,8-dihydroxy-3,7-dimethylocta-2,4-dien-1-yl]-9-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-3-methylpenta-2,4-dienoic acid
-
IC50 (ng/ml): 22.9. Cell death inducibility of osteoclasts (microgram/ml): 6.31
(2E,4E)-6-[(2R,3S,6S,8S,9R)-9-butyl-8-[(1E,3E)-4-carboxy-3-methylbuta-1,3-dien-1-yl]-9-[(3-carboxypropanoyl)oxy]-3-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-4-methylhexa-2,4-dienoic acid
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): not determined
(2E,4S,5S,6E,8E)-10-[(2R,3S,6S,8S,9R)-9-butyl-8-[(1E,3E)-4-carboxy-3-methylbuta-1,3-dien-1-yl]-3-methyl-9-[(methylsulfanyl)methoxy]-1,7-dioxaspiro[5.5]undec-2-yl]-5-hydroxy-4,8-dimethyldeca-2,6,8-trienoic acid
-
IC50 (ng/ml): 497.4. Cell death inducibility of osteoclasts (microgram/ml): 9.6
(2E,4S,5S,6E,8E)-10-[(2R,3S,6S,8S,9R)-9-butyl-8-[(1E,3E)-4-carboxy-3-methylbuta-1,3-dien-1-yl]-9-hydroxy-3-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-5-hydroxy-4,8-dimethyldeca-2,6,8-trienoic acid
-
IC50 (ng/ml): 94.6. Cell death inducibility of osteoclasts (microgram/ml): 11.5
(2E,4S,5S,6E,8E)-10-[(2R,3S,6S,8S,9R)-9-butyl-8-[(1E,3E)-4-carboxy-3-methylbuta-1,3-dien-1-yl]-9-methoxy-3-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-5-hydroxy-4,8-dimethyldeca-2,6,8-trienoic acid
-
IC50 (ng/ml): 57.9. Cell death inducibility of osteoclasts (microgram/ml): not determined
(2E,4S,5S,6E,8E)-10-[(2R,3S,6S,8S,9R)-9-butyl-8-[(1E,3E)-4-carboxy-3-methylbuta-1,3-dien-1-yl]-9-[(3-carboxypropanoyl)oxy]-3-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-4,5-dihydroxy-8-methyldeca-2,6,8-trienoic acid
-
IC50 (ng/ml): 14.4. Cell death inducibility of osteoclasts (microgram/ml): 1.01
(2E,4S,5S,6E,8E)-10-[(2R,3S,6S,8S,9R)-9-butyl-8-[(1E,3E)-4-carboxy-3-methylbuta-1,3-dien-1-yl]-9-[(4-methoxy-4-oxobutanoyl)oxy]-3-methyl-1,7-dioxaspiro[5.5]undec-2-yl]-5-hydroxy-4,8-dimethyldeca-2,6,8-trienoic acid
-
IC50 (ng/ml): 292.8. Cell death inducibility of osteoclasts (microgram/ml): 2.5
(3S)-3-amino-1-bromo-4-methylhexan-2-one
-
labeling reagent
(3S)-3-amino-1-bromo-4-methylpentan-2-one
-
labeling reagent
(3S)-3-amino-1-bromo-4-phenylbutan-2-one
-
labeling reagent
(3S)-3-amino-1-bromoheptan-2-one
-
labeling reagent
1,10-phenanthroline
-
-
2',3'-dialdehyde of tRNAile
-
used to label the binding site for the 3'end of tRNA on the synthetase, incubation of the reagent with IleRS results in a rapid loss of tRNAIle aminoacylation and isoleucine-dependent isotopic ATP-PPi exchange activities
-
2,2'-bipyridyl
-
-
2,3-dideoxy-adenosine-5-[(2S,3S)-2-amino-3-methylpentanoyl]-sulfamate
-
IC50: 0.0064 mM
2,3-Dihydro-5-epireveromycin A
-
IC50 (ng/ml): 58.3. Cell death inducibility of osteoclasts (microgram/ml): 0.82
2,3-Dihydroreveromycin A
-
IC50 (ng/ml): 11.6. Cell death inducibility of osteoclasts (microgram/ml): 0.22
2-deoxy-adenosine-5-[(2S,3S)-2-amino-3-methylpentanoyl]-sulfamate
-
IC50: 0.28 mM
2-fluoro-9-(5-O-phosphono-alpha-L-arabinofuranosyl)-9H-purin-6-amine
-
-
2-fluoroadenosine
-
-
2-iodo-L-isoleucine-NHSO2-AMP
-
highly potent inhibitor, hydrophobic interaction of the 2-substituent of the inhibitor with the adenine binding site of the enzyme
3'-amino-3'-deoxy-N,N-dimethyladenosine
-
-
3'-deoxyadenosine
-
i.e. cordycepin
3-deoxy-adenosine-5-[(2S,3S)-2-amino-3-methylpentanoyl]-sulfamate
-
IC50: 0.035 mM
4,6-dideoxy-4-[[N-(14-methylpentadecanoyl)glycyl]amino]-N-9H-purin-6-ylhexopyranosylamine
-
-
5'-N-[N-(L-isoleucyl)sulfamoyl]adenosine
-
non-hydrolyzable analogue of the reaction intermediate Ile-AMP
5-acetyl-Reveromycin A
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): 0.46
5-Epireveromycin A
-
IC50 (ng/ml): 378.3. Cell death inducibility of osteoclasts (microgram/ml): 21.5
5-methoxy-Reveromycin A
-
IC50 (ng/ml): 374. Cell death inducibility of osteoclasts (microgram/ml): above 15
5-O-succinyl-Spirofungin A
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): 12.4
5-tert-butyl-dimethylsilyl-Reveromycin A
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): 5.9
7-[3-[(2S,3S)-2-amino-3-methyl-1-oxopentyl]amino]propanoyl-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxamide
-
derivative of SB-203207, 10% inhibition at 0.1 mM
7-[4-[(2S,3S)-2-amino-3-methyl-1-oxopentyl]amino]butanoyl-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxamide
7-[[(2S,3S)-2-amino-3-methyl-1-oxopentyl]amino]acetyloxy-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxamide
-
derivative of SB-203207, 17% inhibition at 0.1 mM
7-[[(2S,3S)-2-amino-3-methyl-1-oxopentyl]amino]sulfonylacetyloxy-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxamide
7-[[(2S,3S)-2-amino-3-methyl-1-oxopentyl]amino]sulfonylacetyloxy-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxylic acid methyl ester
7-[[(2S,3S)-2-amino-3-methyl-1-oxopentyl]amino]sulfonylacetyloxy-4,4a,5,6,7,7a-hexahydro-1-methyl-1H-cyclopenta[b]pyridine-3-carboxylic acid methyl ester
7-[[(S)-2-amino-1-oxo-5-thiahexyl]amino]sulfonylacetyloxy-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxamide
7-[[(S)-2-amino-1-oxo-hexyl]amino]sulfonylacetyloxy-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxamide
7-[[(S)-2-amino-3-methyl-1-oxobutyl]amino]sulfonylacetyloxy-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxylic acid methyl ester
7-[[(S)-2-amino-3-methyl-1-oxobutyl]amino]sulfonylacetyloxy-4,4a,5,6,7,7a-hexahydro-1-methyl-1H-cyclopenta[b]pyridine-3-carboxylic acid methyl ester
7-[[(S)-2-amino-3-methyl-1-oxopentyl]amino]sulfonylacetyloxy-4,4a,5,6,7,7a-hexahydro-1-methyl-1H-cyclopenta[b]pyridine-3-carboxylic acid methyl ester
7-[[(S)-2-amino-4-methyl-1-oxopentyl]amino]sulfonylacetyloxy-2,4a,5,6,7,7a-hexahydro-2-methyl-1H-cyclopenta[c]pyridine-4-carboxylic acid methyl ester
-
derivative of SB-203207, 14% inhibition at 0.1 mM
8-Aminoadenosine
-
-
8-azidoadenosine 5'-triphosphate
-
-
9-(3-deoxy-alpha-L-threo-pentofuranosyl)-9H-purin-6-amine
-
-
9-(5-O-acetyl-alpha-L-arabinofuranosyl)-9H-purin-6-amine
-
-
9-(alpha-D-arabinofuranosyl)-9H-purin-6-amine
-
competitive inhibition
9-(alpha-L-arabinofuranosyl)-9H-purin-6-amine
-
-
9-(alpha-L-arabinofuranosyl)-N-(3-methylbut-2-en-1-yl)-9H-purin-6-amine
-
-
9-(beta-D-arabinofuranosyl)-9H-purin-6-amine
-
-
9-[5-deoxy-5-[(methylsulfonyl)amino]-alpha-L-arabinofuranosyl]-9H-purin-6-amine
-
-
9-[5-O-(dimethoxyphosphoryl)-alpha-L-arabinofuranosyl]-9H-purin-6-amine
-
-
9-[5-O-[tert-butyl(dimethyl)silyl]-alpha-L-arabinofuranosyl]-9H-purin-6-amine
-
-
adenosine-5-[(2S,3S)-2-amino-3-methylpentanoyl]-sulfamate
-
IC50: 0.000265 mM
Chymotrypsin
-
proteolytic inactivation patterns, bound Ile-AMP or inhibitors isoleucinol adenylate and pseudomonic acid protect, 50fold higher concentration is needed for digestion of Ile-AMP-enzyme complex than for the free enzyme at 37C
-
diethyl dicarbonate
-
-
diphosphate
-
partly inhibits the binding of tRNA
ester analogues of isoleucyl adenylate
-
with or without cyclic substitutents at the adenine moiety
ethyl monate-A
-
-
Furanomycin
-
-
hydroxamate analogues of isoleucyl adenylate
-
with or without cyclic substitutents at the adenine moiety
Ile-NHSO2-AMP
-
non-hydrolyzable reaction intermediate analogue, slow-tight binding, competitive and inhibition mechanism, reversible
Ile-ol-AMP
-
-
isoleucinol adenylate
-
determination of binding structures, bound inhibitor protects against proteolytic inactivation by trypsin or chymotrypsin and specifically alters the proteolytic cleavage pattern
isoleucinyl adenylate
-
i.e. Ile-ol-AMP, nonhydrolyzable reaction intermediate analogue, competitive with respect to both ATP and Ile
isoleucinyl-adenylate
-
i.e. Ile-ol-AMP, non-hydrolyzable reaction intermediate analogue, slow-tight binding, competitive and inhibition mechanism, reversible
isoleucyl isovanilloids
-
e.g. the isovanillic hydroxamate and amide analogue
isoleucyl sulfamate analogues
-
-
isoleucyl vanilloids
-
e.g. the vanillic hydroxamate with a phenolic hydoxyl at the para-position
isoleucyl-N'-adenosyl-N'-hydroxy sulfamide
-
-
isoleucyl-N'-adenosyloxy sulfamide
-
-
Mg2+
-
in presence of 50 mM K+ and in absence of polyamines, the optimal Mg2+ concentration for Ile-tRNA formation is 1 mM, an increase in Mg2+ concentration markedly inhibits
muciproin
mupirocin
N8,N8-dimethyl-9-pentofuranosyl-9H-purine-6,8-diamine
-
-
NSC70422
-
-
Pseudomonic acid
purineriboside
-
i.e. nebularin
pyridoxal 5'-diphospho-5'-adenosine
-
affinity labeling reagent for the ATP-binding site, incubation of the reagent with IleRS results in a rapid loss of tRNAIle aminoacylation and isoleucine-dependent isotopic ATP-PPi exchange activities
Reveromycin A
-
IC50 (ng/ml): 2.95. Cell death inducibility of osteoclasts (microgram/ml): 0.06
Reveromycin A 1-methyl ester
-
IC50 (ng/ml): 211. Cell death inducibility of osteoclasts (microgram/ml): 2
Reveromycin A 24-methyl ester
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): 3.1
Reveromycin B
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): above 15
SB-203207
SB-205952
spermine
-
catalyzes ATP-diphosphate exchange, no inhibition of specific aminoacylation of tRNAIle
Spirofungin A
-
IC50 (ng/ml): 564.5. Cell death inducibility of osteoclasts (microgram/ml): above 30
Spirofungin B
-
IC50 (ng/ml): value above 1000. Cell death inducibility of osteoclasts (microgram/ml): above 30
thiaisoleucine
-
directly competes with isoleucine for a target, irreversible inhibition, inhibits ring-stage parasites in development
tRNA
-
partly inhibits the binding of diphosphate
Trypsin
-
proteolytic inactivation patterns, bound Ile-AMP or inhibitors isoleucinol adenylate and pseudomonic acid protect, 50fold higher concentration is needed for digestion of Ile-AMP-enzyme complex than for the free enzyme at 37C
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
spermidine
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00028 - 4.4
ATP
0.0036 - 1.3
Ile
0.00000004 - 52.8
L-isoleucine
0.0001 - 0.0021
tRNAIle
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.29 - 80.4
ATP
27.7 - 104
isoleucine
0.18 - 18
L-isoleucine
3.1
tRNAIle
Escherichia coli
-
wild-type enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2 - 1.49
ATP
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000027
9-(alpha-D-arabinofuranosyl)-9H-purin-6-amine
-
pH and temperature not specified in the publication
0.00004
ethyl monate-A
-
pH 7.9, 22C
0.000003
Ile-NHSO2-AMP
-
pH 7.9, 22C
0.00003
Ile-ol-AMP
0.00009
isoleucinyl-adenylate
-
pH 7.9, 22C
0.00025 - 0.0023
muciproin
0.0000027
NSC70422
-
pH and temperature not specified in the publication
0.000001 - 0.00002
Pseudomonic acid
0.000045
SB-205952
-
pH 7.9, 22C
additional information
additional information
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0064
2,3-dideoxy-adenosine-5-[(2S,3S)-2-amino-3-methylpentanoyl]-sulfamate
Escherichia coli
-
IC50: 0.0064 mM
0.28
2-deoxy-adenosine-5-[(2S,3S)-2-amino-3-methylpentanoyl]-sulfamate
Escherichia coli
-
IC50: 0.28 mM
0.035
3-deoxy-adenosine-5-[(2S,3S)-2-amino-3-methylpentanoyl]-sulfamate
Escherichia coli
-
IC50: 0.035 mM
0.0000141
9-(alpha-D-arabinofuranosyl)-9H-purin-6-amine
Trypanosoma brucei
-
pH and temperature not specified in the publication
0.000265
adenosine-5-[(2S,3S)-2-amino-3-methylpentanoyl]-sulfamate
Escherichia coli
-
IC50: 0.000265 mM
0.0000141
NSC70422
Trypanosoma brucei
-
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00084
-
-
0.06
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.3
activity assay
additional information
-
pH-dependence of enzyme-substrate complex formation
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.8
-
about 30% of maximal activity at pH 6.0 and 8.8
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
formation of isoleucyl-tRNA
62
-
isoleucine-tRNA formation with E. coli tRNA
65
-
isoleucine-dependent ATP-diphosphate exchange reaction
75
-
isoleucine tRNA formation with Thermus thermophilus tRNA
80
-
isoleucine dependent ATP-diphosphate exchange
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
cell line LCC-RK1
Manually annotated by BRENDA team
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cell line CRL-1781
Manually annotated by BRENDA team
additional information
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the cytoplasmic isozyme gene is expressed during the erythrocytic stages and is localized in the proper compartment for the bulk of protein synthesis, while the apicoplast isozyme is expressed at low rates in the erythrocytic stage and gets more abundant in later developmental stages, overview
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
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two IRS isozymes localize to distinct compartments in the parasite
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Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
102000
-
sedimentation equilibrium measurement
115000
200000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
analysis of the crystal structure with bound muciproin, PDB ID: 1FFY
purified recombinant enzyme cocrystallized with Escherichia coli tRNAIle and inhibitor muciproin, formation of the socalled editing complex, X-ray diffraction structure determination at 2.2 A resolution, and structure analysis
crystallization of CP1 domain alone or in complex with L-valine, X-ray structure determination at 1.8 and 2.0 A resolution respectively, and analysis
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crystallization of the 5'-N-[N-(L-isoleucyl)sulfamoyl]adenosine-enzyme complex, and the muciproin-enzyme complex by preparation of enzyme crystals and soaking of the crystals in 0.1 mM 5'-N-[N-(L-isoleucyl)sulfamoyl]adenosine solution, and 1 mM muciproin solution, respectively, X-ray diffraction structure determination at 2.5 A resolution, and analysis
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IleRS editing domain complexed with the substrate analogues in the pre and post-transfer modes, X-ray diffraction structure determination and analysis at 1.7 A resolution
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
77
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half-life is 15 min, inactivation is completely suppressed by addition of either E. coli or Thermus thermophilus tRNA
additional information
-
isoleucine, isoleucine + ATP and tRNA + Mg2+ protect against heat inactivation, some protection by high concentrations of valine
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, 50% glycerol
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-20C, 50% glycerol
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme, and recombinant enzyme from Escherichia coli, amino acid sequence determination
recombinant His-tagged IleRS from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
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recombinant His-tagged wild-type and mutants enzymes from Escherichia coli by nickel affinity chromatography
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using M2-FLAG resin
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
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expression in Escherichia coli
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expression of His-tagged wild-type and mutants enzymes in Escherichia coli
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gene ileS, quantitative real-time PCR expression analysis
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gene ileS2, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strains DH5alpha and Ts331 mediates resistance against pseudomonic acid, complementation of the ileS-deficient strain Ts331
genotyping of thiaisoleucine-resistant enzymes, expression of GFP-tagged cytoplasmic isozyme in erythrocytic stage parasite
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into the vector pET3D-FLAG for expression in Escherichia coli BL21DE3 cells
overexpression of His-tagged IleRS in Escherichia coli strain BL21 (DE3)
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with the pCR2.1-TOPO TA cloning kit for sequencing
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
AIleRS
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mutant enzymes IleRS(C922S) and AIleRS with replacement of Cys922 through Ala939 with a 33 amino acid peptide unable to bind zinc. Mutant enzymes have altered zinc binding and aminoacylation activity
D342A
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site-directed mutagenesis, the IleRS CP1 domain mutant is unable to deacylate misacylated tRNA even at high enzyme concentrations
IleRS(C922S)
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mutant enzymes IleRS(C922S) and AIleRS with replacement of Cys922 through Ala939 with a 33 amino acid peptide unable to bind zinc. Mutant enzymes have altered zinc binding and aminoacylation activity
T243R
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site-directed mutagenesis, the mutant retains tRNA-independent editing at a level identical to the WT enzyme and shows increased ATP hydrolysis compared to the wild-type enzyme
T243R/D342A
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site-directed mutagenesis, the IleRS CP1 domain mutant is unable to deacylate misacylated tRNA even at high enzyme concentrations
L810F
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naturally occuring mutation in the cytoplasmic IleRS responsible for thiaisoleucine-resistance in the parasite, phenotype, overview
P184T
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naturally occuring mutation that restores fitness in mupirocin resistant strains
Q420H
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naturally occuring mutation that restores fitness in mupirocin resistant strains
F227L
the naturally occuring mutation affects the muciprocin binding
H581L/L583H
site-directed mutagenesis, slightly reduced enzyme activity
K226T
the naturally occuring mutation affects the muciprocin binding
P187F
the naturally occuring mutation affects the muciprocin binding
Q612H
the naturally occuring mutation is involved in stabilizing the conformation of the catalytic loop containing the KMSKS motif
V588F
the naturally occuring mutation affects the Rossman fold and leads to low-level mupirocin resistance
V767D
the naturally occuring mutation affects the muciprocin binding
H319A
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site-directed mutagenesis, Thr233 and His319 recognize the substrate valine side-chain, regardless of the valine side-chain rotation, and reject the isoleucine side-chain, but the mutant shows detectable editing activities against the cognate isoleucine, mechanism, overview
T223A
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site-directed mutagenesis, Thr233 and His319 recognize the substrate valine side-chain, regardless of the valine side-chain rotation, and reject the isoleucine side-chain, but the mutant shows detectable editing activities against the cognate isoleucine, mechanism, overview
W227A
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site-directed mutagenesis, both editing activities of the mutant are reduced compared to the wild-type enzyme
W227F
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site-directed mutagenesis, the mutant shows editing activities which are unaltered compared to the wild-type enzyme
W227H
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site-directed mutagenesis, both editing activities of the mutant are reduced compared to the wild-type enzyme
W227L
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site-directed mutagenesis, both editing activities of the mutant are reduced compared to the wild-type enzyme
W227V
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site-directed mutagenesis, both editing activities of the mutant are reduced compared to the wild-type enzyme
W227Y
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site-directed mutagenesis, the mutant shows editing activities which are unaltered compared to the wild-type enzyme
D539A/W541A
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catalytically inactive
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
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isoleucine utilization is an essential pathway that can be targeted for antimalarial drug development
medicine
molecular biology
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enzyme is a target for receptor-guided inhibitor design
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