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Information on EC 6.1.1.9 - valine-tRNA ligase and Organism(s) Escherichia coli and UniProt Accession P07118
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6.1.1.9 valine-tRNA ligaseSpecify your search results
Word Map
- 6.1.1.9
- synthetases
- aminoacylation
- aminoacyl-trna
-
valylation
-
isoleucyl-trna
-
anticodon
- ilers
-
turnip
-
noncognate
- leucyl-trna
-
mischarged
-
trna-like
-
isoleucyl
-
misacylated
-
misactivated
-
post-transfer
-
l-shaped
-
aarss
- phenylalanyl-trna
-
threonylation
- trnaile
- cysrs
-
atp-ppi
- medicine
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
Synonyms
valyl-trna synthetase, valrs, glp-4, val-trna synthetase, valyl-transfer rna synthetase, osvalrs2, valine-trna ligase, valyl-trna ligase, valyl transfer ribonucleic acid synthetase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
G7a
Synthetase, valyl-transfer ribonucleate
Valine transfer ribonucleate ligase
Valine translase
Valine--tRNA ligase
ValRS
Valyl transfer ribonucleic acid synthetase
Valyl-transfer ribonucleate synthetase
Valyl-transfer RNA synthetase
Valyl-tRNA ligase
Valyl-tRNA synthetase
G7a
-
-
-
-
Synthetase, valyl-transfer ribonucleate
-
-
-
-
Valine transfer ribonucleate ligase
-
-
-
-
Valine translase
-
-
-
-
Valine--tRNA ligase
-
-
-
-
ValRS
-
-
-
-
Valyl transfer ribonucleic acid synthetase
-
-
-
-
Valyl-transfer ribonucleate synthetase
-
-
-
-
Valyl-transfer RNA synthetase
-
-
-
-
Valyl-tRNA ligase
-
-
-
-
Valyl-tRNA synthetase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-valine + tRNAVal = AMP + diphosphate + L-valyl-tRNAVal
Lys593 and Lys554 are in the active site binding valine or threonine, part of the 554KMSKS558 consensus sequence, mechanism, determination of diverse amino acid residues of the enzyme molecule involved in substrate binding, Lys277 plays a crucial role in the fidelity of tRNA aminoacylation by the enzyme, nucleophilic attack of misacylated tRNA in the diting site of the enzyme
-
ATP + L-valine + tRNAVal = AMP + diphosphate + L-valyl-tRNAVal
substrate binding mechanism, bases A73 and C75 are involved in the tRNA substrate recognition and editing reaction
-
ATP + L-valine + tRNAVal = AMP + diphosphate + L-valyl-tRNAVal
tRNAVal 3'-end, and a purine at position 76, is crucial for substrate editing mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
esterification
Aminoacylation
esterification
-
-
-
-
Aminoacylation
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
L-valine:tRNAVal ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY
9023-47-6
-
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
ATP + DL-2-amino-3-chlorobutyrate + tRNAVal
AMP + diphosphate + DL-2-amino-3-chlorobutyryl-tRNAVal
-
as effective as valine
-
-
?
ATP + DL-2-aminobutyrate + tRNAVal
AMP + diphosphate + DL-2-aminobutyryl-tRNAVal
-
30% of the activity with valine
-
-
?
ATP + DL-allo-2-amino-3-chlorobutyrate + tRNAVal
AMP + diphosphate + DL-allo-2-amino-3-chlorobutyryl-tRNAVal
-
15% of the activity with valine
-
-
?
ATP + L-isoleucine + tRNAVal
AMP + diphosphate + L-isoleucyl-tRNAVal
-
very low activity
-
?
ATP + L-threonine + tRNAVal
?
-
1 step performance of an originally two-step reaction, the enzyme can hardly differentiate between the cognate amino acid valine and others, especially threonine, to minimize misaminoacetylation the enzyme performs a proofreading, socalled editing reaction at a second active site, which is dependent on the presence of cognate tRNAVal whose 3'-end is involved in the editing reaction, a majority of editing by the enzyme entails prior charging of the tRNA, misacylated tRNA is a transient intermediate in the editing reaction
-
?
ATP + L-valine + tRNAVal,3' 2-aminopurine
AMP + diphosphate + L-valyl-tRNAVal, 3' 2-aminopurine
-
mutant tRNAVal with 3'-terminal 2-aminopurine base analogue substitution
-
?
ATP + L-valine + tRNAVal,3' inosine
AMP + diphosphate + L-valyl-tRNAVal, 3' inosine
-
mutant tRNAVal with 3'-terminal inosine base analogue substitution
-
?
ATP + L-valine + tRNAVal,3' isoguanosine
AMP + diphosphate + L-valyl-tRNAVal, 3' isoguanosine
-
mutant tRNAVal with 3'-terminal isoguanosine base analogue substitution
-
?
ATP + L-valine + tRNAVal,3' purine riboside
AMP + diphosphate + L-valyl-tRNAVal, 3' purine riboside
-
mutant tRNAVal with 3'-terminal purine riboside base analogue substitution
-
?
L-threonyl-tRNAVal
L-threonine + tRNAVal
-
substrate bound to the enzyme, intermediate in the posttransfer editing reaction
-
ir
L-threonyl-tRNAVal-AMP
L-threonine + tRNAVal + AMP
-
substrate bound to the enzyme, intermediate in the pretransfer editing reaction
-
ir
ATP + L-valine + tRNAVal
AMP + diphosphate + L-valyl-tRNAVal
charging of tRNAVal by ValS occurs in two steps: firstly, valine is activated with ATP to form Val-AMP leading to the release of pyrophosphate, and secondly, the valine is then transferred to the tRNAVal to form Val-tRNAVal, with a concomitant release of AMP
-
-
?
ATP + L-threonine + tRNAVal
AMP + diphosphate + L-threonyl-tRNAVal
-
28% of the activity with valine
-
-
?
ATP + L-threonine + tRNAVal
AMP + diphosphate + L-threonyl-tRNAVal
-
noncognate isosteric substrate, low activity, posttransfer editing occurs
-
ir
ATP + L-valine + tRNAVal
AMP + diphosphate + L-valyl-tRNAVal
-
-
-
-
?
ATP + L-valine + tRNAVal
AMP + diphosphate + L-valyl-tRNAVal
-
-
-
ir
ATP + L-valine + tRNAVal
AMP + diphosphate + L-valyl-tRNAVal
-
enzyme contains 2 tRNA binding sites involved in aminoacetylation and editing reactions, misacetylated tRNAVal is edited by the enzyme to avoid accumulation, the 3'-end of the tRNA is involved
-
?
ATP + L-valine + tRNAVal
AMP + diphosphate + L-valyl-tRNAVal
-
two-step reaction, covalent valylation of the enzyme
-
ir
ATP + L-valine + tRNAVal
AMP + diphosphate + L-valyl-tRNAVal
-
two-step reaction, the enzyme can hardly differentiate between the cognate amino acid valine and others, especially threonine, to minimize misaminoacetylation the enzyme performs a proofreading, socalled editing reaction at a second active site, which is dependent on the presence of cognate tRNAVal whose 3'-end is involved in the editing reaction, a majority of editing by the enzyme entails prior charging of the tRNA, misacylated tRNA is a transient intermediate in the editing reaction
-
?
additional information
?
-
-
valine-dependent ATP-diphosphate exchange: valine + ATP + enzyme /Val-AMP-enzyme + diphosphate
-
-
?
additional information
?
-
-
catalytic activity with L-valine and diverse tRNA analogues with functional group substitutions at the terminal nucleoside of the 3'-end, some of which stimulate the editing reaction, overview
-
?
additional information
?
-
-
substrate specificity with diverse Escherichia coli tRNAs and tRNA mutants in editing and aminoacylation reaction, overview
-
?
additional information
?
-
-
the enzyme also performs the ATP-diphosphate exchange reaction, labeling of the enzyme by methionyladenylate, determination of lysine residues involved in binding
-
?
additional information
?
-
-
tRNAVal variants in position 76, i.e. U76, C76, G76 activate the editing activity, misactivation of threonine, alanine, serine, cysteine, alpha-aminobutyrate, and to a low extent of norvaline activates the editing reaction at different rates, overview, the enzyme is unable to deacylate misacylated tRNAVal terminating in 3'-pyrimidines but does deacylate mischarged tRNAVal terminating in adenosine or guanosine, no misactivation of methionine, leucine, glycine, glutamic acid, lysine, tyrosine, and phenylalanine
-
?
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
ATP + L-valine + tRNAVal
AMP + diphosphate + L-valyl-tRNAVal
-
-
-
ir
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-triphospho-5'-adenosine
-
aminoacylation and ATP-diphosphate exchange abolished
additional information
-
incorporation of 2.1 mol L-valine per mol of enzyme leads to inactivation
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
stimulatory effects of diverse tRNA variants on the editing activity, overview
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.3
L-threonine
-
ATP-diphosphate exchange reaction, native wild-type enzyme, pH 7.5, 25°C
additional information
additional information
-
steady-state parameters for tRNA-independent pre-transfer editing by ValRS and its mutants determined by varying concentrations of noncognate threonine, overview
-
0.11
ATP
-
ATP-diphosphate exchange reaction, His-tagged wild-type and mutant enzymes, pH 7.5, 25°C
0.0019
L-valine
-
aminoacylation reaction, His-tagged K277A mutant enzyme, pH 7.5, 25°C
0.002
L-valine
-
with mutant tRNAVal with 3'-terminal isoguanosine base analogue, pH 7.5, 37°C
0.0048
L-valine
-
with mutant tRNAVal with 3'-terminal inosine base analogue, pH 7.5, 37°C
0.005
L-valine
-
with mutant tRNAVal with 3'-terminal 2-aminopurine base analogue, pH 7.5, 37°C
0.0088
L-valine
-
with mutant tRNAVal with 3'-terminal purine riboside base analogue, pH 7.5, 37°C
0.047
L-valine
-
aminoacylation reaction, His-tagged wild-type enzyme, pH 7.5, 25°C
0.062
L-valine
-
ATP-diphosphate exchange reaction, native wild-type enzyme, pH 7.5, 25°C
0.07
L-valine
-
ATP-diphosphate exchange reaction, His-tagged wild-type enzyme, pH 7.5, 25°C
0.072
L-valine
-
ATP-diphosphate exchange reaction, His-tagged K277A mutant enzyme, pH 7.5, 25°C
0.00008
tRNAVal
-
aminoacylation reaction with L-valine, wild-type enzyme, pH 7.5, 25°C
0.0001
tRNAVal
-
aminoacylation reaction with L-valine, His-tagged wild-type enzyme, pH 7.5, 25°C
0.0001
tRNAVal
-
aminoacylation reaction with L-valine, K277A mutant enzyme, pH 7.5, 25°C
0.00026
tRNAVal
-
aminoacylation reaction with L-valine, His-tagged K277A mutant enzyme, pH 7.5, 25°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
rate of hydrolysis of Thr-tRNAVal and Val-tRNAVal by wild-type and mutant enzyme, incorporation of amino acids valine and threonine into the enzyme
-
50
ATP
-
ATP-diphosphate exchange reaction, His-tagged wild-type enzyme, pH 7.5, 25°C
57
ATP
-
ATP-diphosphate exchange reaction, His-tagged K277A mutant enzyme, pH 7.5, 25°C
0.064
L-valine
-
aminoacylation reaction, His-tagged K277A mutant enzyme, pH 7.5, 25°C
0.68
L-valine
-
with mutant tRNAVal with 3'-terminal 2-aminopurine base analogue, pH 7.5, 37°C
0.99
L-valine
-
with mutant tRNAVal with 3'-terminal isoguanosine base analogue, pH 7.5, 37°C
1.06
L-valine
-
with mutant tRNAVal with 3'-terminal inosine base analogue, pH 7.5, 37°C
1.5
L-valine
-
aminoacylation reaction, His-tagged wild-type enzyme, pH 7.5, 25°C
6.08
L-valine
-
with mutant tRNAVal with 3'-terminal 2-aminopurine base analogue, pH 7.5, 37°C
6.08
L-valine
-
with mutant tRNAVal with 3'-terminal inosine base analogue, pH 7.5, 37°C
6.08
L-valine
-
with mutant tRNAVal with 3'-terminal isoguanosine base analogue, pH 7.5, 37°C
14.4
L-valine
-
with mutant tRNAVal with 3'-terminal purine riboside base analogue, pH 7.5, 37°C
1.5
tRNAVal
-
aminoacylation reaction with L-valine, K277A mutant enzyme, pH 7.5, 25°C
1.7
tRNAVal
-
aminoacylation reaction with L-valine, wild-type enzyme, pH 7.5, 25°C
50
tRNAVal
-
ATP-diphosphate exchange reaction, K277A mutant enzyme, pH 7.5, 25°C
51
tRNAVal
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.5, 25°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
catalytic activity with L-valine and diverse tRNA analogues with functional group substitutions at the terminal nucleoside of the 3'-end, overview
additional information
-
in ValRS transfer to tRNA is 200fold faster than hydrolysis
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
the proline triplet in ValS, the tRNA synthetase that charges tRNAVal with valine, is the only single polyproline stretch that is invariant across all domains of life. The critical role of the proline triplet for ValS activity may explain why bacterial cells co-evolved the EF-P rescue system. Nature has evolved not only specialized translation factors to overcome stalling at polyproline stretches, but also evolved independent sets of modification enzymes to activate these factors. This in itself implies that the benefits of retaining polyproline stretches significantly outweigh the cost of implementing and maintaining the EF-P and a/eIF5A rescue systems
physiological function
the proline triplet located in the active site of ValS is important for efficient charging of tRNAVal with valine preventing formation of mischarged Thr-tRNAVal, as well as for efficient growth of Escherichia coli in vivo. The critical role of the proline triplet for ValS activity may explain why bacterial cells co-evolved the EF-P rescue system
physiological function
-
hydrolytic editing activities are present in aminoacyl-tRNA synthetases possessing reduced amino acid discrimination in the synthetic reactions. Post-transfer hydrolysis of misacylated tRNA in class I editing enzymes, e.g. ValRS, occurs in a spatially separate domain inserted into the catalytic Rossmann fold, location and mechanisms of pre-transfer hydrolysis of misactivated amino acids, overview. The rates of amino acid transfer to tRNA are similar for cognate and noncognate aminoacyl-adenylates. Editing by ValRS occurs nearly exclusively by post-transfer hydrolysis in the editing domain
additional information
expression of ValS is strictly dependent on the presence of active elongation factor P (EF-P) in vivo and in vitro
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
nucleotide sequence of the Escherichia coli valyl-tRNA synthetase gene
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D286A
-
site-directed mutagenesis, the ValRS CP1 domain mutant is unable to deacylate misacylated tRNA even at high enzyme concentrations
K277A
-
site-directed mutagenesis, mutant exhibits a reduced posttransfer editing activity compared to the wild-type, also the specificiy of the editing reaction is modulated, the mutant hydrolyzes the correctly formed Val-tRNAVal, increased sensitivity to Mg2+, high concentrations inactivate
K277P
-
site-directed mutagenesis, the ValRS CP1 domain mutant is unable to deacylate misacylated tRNA even at high enzyme concentrations
K277P/D286A
-
site-directed mutagenesis, the ValRS CP1 domain mutant is unable to deacylate misacylated tRNA even at high enzyme concentrations
additional information
additional information
generation of three single ValS Pro-to-Gly mutants, ValS-GPP, ValS-PGP, and ValS-PPG, as well as a triple PPP-to-GGG mutant (ValS-GGG). The ability of the ValS mutants to charge tRNAVal with [14C]valine is assessed and compared to wild-type ValS. All ValS mutants are less efficient than the wild-type, the ValS-PPG and ValS-GGG mutants are completely devoid of activity, whereas the ValS-PGP and ValS-GPP mutants retain some activity but at lower levels than the wild-type enzyme
additional information
-
substitution of base 75 of C by U or A leads to effective stimulation of the editing activity at lower rates, substitution with G leads to reduction of the editing activity by 95%, mutational exchange of the discriminator base A73 to U73 or C73 does not alter enzyme activity but exchange to G73 stimulates editing activity of the catalytically inactive mutant
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged ValRS from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene valS, expression of His-tagged wild-type and mutant enzymes in strain JM101Tr
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression of ValS in vivo and in vitro requires the elongation factor P (EF-P), the downregulation of ValS in the absence of active EF-P is specific, the lack of EF-P leads to a strong downregulation of ValS. The lack of YfcM (DELTAyfcM), which hydroxylates the lysinylated EF-P, does not lead to downregulation of ValS. ValS expression is also dependent on EF-P in vitro
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Hountondji, C.; Schmitter, J.M.; Fukui, T.; Tagaya, M.; Blanquet, S.
Affinity labeling of aminoacyl-tRNA synthetases with adenosine triphosphopyridoxal. Probing the Lys-Met-Ser-Lys-Ser signature sequence as the ATP-binding site in Escherichia coli methionyl- and valyl-tRNA synthetases
Biochemistry
29
11266-11273
1990
Escherichia coli
Bergmann, F.H.; Berg, P.; Dieckmann, M.
The Enzymic synthesis of amino acyl derivatives of ribonucleic acid. II. The preparation of leucyl-, valyl-, isoleucyl-, and methionyl ribonucleic acid synthetases from Escherichia coli
J. Biol. Chem.
236
1735-1740
1961
Escherichia coli
-
Paradies, H.H.
Isolation and crystallization of valyl-tRNA synthetase from E. coli MRE 600
J. Biochem.
76
655-659
1974
Escherichia coli, Escherichia coli MRE 600
Hrtlein, M.; Frank, R.; Madern, D.
Nucleotide sequence of Escherichia coli valyl-tRNA synthetase gene valS
Nucleic Acids Res.
15
9081-9082
1987
Escherichia coli
Heck, J.D.; Hatfield, G.W.
Valyl-tRNA synthetase gene of Escherichia coli K12. Primary structure and homology within a family of aminoacyl-tRNA synthetases
J. Biol. Chem.
263
868-877
1988
Escherichia coli
Tardif, K.D.; Liu, M.; Vitseva, O.; Hou, Y.M.; Horowitz, J.
Misacylation and editing by Escherichia coli valyl-tRNA synthetase: evidence for two tRNA binding sites
Biochemistry
40
8118-8125
2001
Escherichia coli
Hountondji, C.; Lazennec, C.; Beauvallet, C.; Dessen, P.; Pernollet, J.C.; Plateau, P.; Blanquet, S.
Crucial role of conserved lysine 277 in the fidelity of tRNA aminoacylation by Escherichia coli valyl-tRNA synthetase
Biochemistry
41
14856-14865
2002
Escherichia coli
Tardif, K.D.; Horowitz, J.
Transfer RNA determinants for translational editing by Escherichia coli valyl-tRNA synthetase
Nucleic Acids Res.
30
2538-2545
2002
Escherichia coli
Tardif, K.D.; Horowitz, J.
Functional group recognition at the aminoacylation and editing sites of E. coli valyl-tRNA synthetase
RNA
10
493-503
2004
Escherichia coli
Dulic, M.; Cvetesic, N.; Perona, J.J.; Gruic-Sovulj, I.
Partitioning of tRNA-dependent editing between pre- and post-transfer pathways in class I aminoacyl-tRNA synthetases
J. Biol. Chem.
285
23799-23809
2010
Escherichia coli
Cvetesic, N.; Perona, J.J.; Gruic-Sovulj, I.
Kinetic partitioning between synthetic and editing pathways in class I aminoacyl-tRNA synthetases occurs at both pre-transfer and post-transfer hydrolytic steps
J. Biol. Chem.
287
25381-25394
2012
Escherichia coli
Starosta, A.L.; Lassak, J.; Peil, L.; Atkinson, G.C.; Woolstenhulme, C.J.; Virumaee, K.; Buskirk, A.; Tenson, T.; Remme, J.; Jung, K.; Wilson, D.N.
A conserved proline triplet in Val-tRNA synthetase and the origin of elongation factor P
Cell Rep.
9
476-483
2014
Escherichia coli (P07118)
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