Information on EC 6.1.1.3 - Threonine-tRNA ligase

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

EC NUMBER
COMMENTARY
6.1.1.3
-
RECOMMENDED NAME
GeneOntology No.
Threonine-tRNA ligase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
; bi uni uni bi ping-pong mechanism
-
-
-
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
induced fit, Trp434 is involved in conformational changes during substrate binding
-
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
functional mechanism and substrate recognition, editing model, 2 separate active sites for substrate binding, binding of tRNA is more specific than the binding of the amino acid
-
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
mechanism, active site structure, and substrate recognition method
-
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
substrate binding and mechanism
Q8NW68, -
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
the catalytic domain is located at the C-terminus of the enzyme
-
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
active site structure
-
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
determination of substrate binding sites, catalytic sites, and catalytic mechanism from structure modeling
-
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
reaction mechanism analysis by QM/MM and MM modeling of full length and truncated enzyme, molecular mechanism simulation
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Aminoacylation
-
-
-
-
Aminoacylation
Q8NW68, -
-
Aminoacylation
Q9YDW0, Q9YFY3
-
Aminoacylation
-
-
Aminoacylation
-
-
Aminoacylation
Q9YDW0, Q9YFY3
;
Aminoacylation
-
-
esterification
-
-
-
-
esterification
Q8NW68, -
-
esterification
Q9YDW0, Q9YFY3
-
esterification
-
-
PATHWAY
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
tRNA charging
-
SYSTEMATIC NAME
IUBMB Comments
L-Threonine:tRNAThr ligase (AMP-forming)
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
ApThrRS-1
-
protein contains the catalytic domain of ThrRS
ApThrRS-1
Q9YDW0
-
ApThrRS-2
-
protein contains the editing domain of ThrRS
ApThrRS-2
Q9YFY3
-
SfThrRS-1
-
protein contains the catalytic domain of ThrRS
SfThrRS-2
-
protein contains the editing domain of ThrRS
Synthetase, threonyl-transfer ribonucleate
-
-
-
-
Synthetase, threonyl-transfer ribonucleate
Q9YFY3
-
Synthetase, threonyl-transfer ribonucleate
-
-
Synthetase, threonyl-transfer ribonucleate
-
-
Synthetase, threonyl-transfer ribonucleate
-
-
Synthetase, threonyl-transfer ribonucleate
-
-
Synthetase, threonyl-transfer ribonucleate
-
-
Thr-tRNA synthetase
-
-
Thr-tRNA synthetase
-
-
Threonine translase
-
-
-
-
Threonine translase
Q9YFY3
-
Threonine translase
-
-
Threonine translase
-
-
Threonine translase
-
-
Threonine translase
-
-
Threonine--tRNA ligase
-
-
-
-
Threonine--tRNA ligase
Q9YFY3
-
Threonine--tRNA ligase
-
-
Threonine--tRNA ligase
-
-
Threonine--tRNA ligase
-
-
Threonine--tRNA ligase
-
-
Threonine-transfer ribonucleate synthetase
-
-
-
-
Threonine-transfer ribonucleate synthetase
Q9YFY3
-
Threonine-transfer ribonucleate synthetase
-
-
Threonine-transfer ribonucleate synthetase
-
-
Threonine-transfer ribonucleate synthetase
-
-
Threonine-transfer ribonucleate synthetase
-
-
Threonine-transfer ribonucleate synthetase
-
-
Threonyl-ribonucleic synthetase
-
-
-
-
Threonyl-ribonucleic synthetase
Q9YFY3
-
Threonyl-ribonucleic synthetase
-
-
Threonyl-ribonucleic synthetase
-
-
Threonyl-ribonucleic synthetase
-
-
Threonyl-ribonucleic synthetase
-
-
Threonyl-ribonucleic synthetase
-
-
Threonyl-transfer ribonucleate synthetase
-
-
-
-
Threonyl-transfer ribonucleate synthetase
Q9YFY3
-
Threonyl-transfer ribonucleate synthetase
-
-
Threonyl-transfer ribonucleate synthetase
-
-
Threonyl-transfer ribonucleate synthetase
-
-
Threonyl-transfer ribonucleate synthetase
-
-
Threonyl-transfer ribonucleate synthetase
-
-
Threonyl-transfer ribonucleic acid synthetase
-
-
-
-
Threonyl-transfer ribonucleic acid synthetase
Q9YFY3
-
Threonyl-transfer ribonucleic acid synthetase
-
-
Threonyl-transfer ribonucleic acid synthetase
-
-
Threonyl-transfer ribonucleic acid synthetase
-
-
Threonyl-transfer ribonucleic acid synthetase
-
-
Threonyl-transfer ribonucleic acid synthetase
-
-
Threonyl-transfer RNA synthetase
-
-
-
-
Threonyl-transfer RNA synthetase
Q9YFY3
-
Threonyl-transfer RNA synthetase
-
-
Threonyl-transfer RNA synthetase
-
-
Threonyl-transfer RNA synthetase
-
-
Threonyl-transfer RNA synthetase
-
-
Threonyl-transfer RNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
Q9YDW0
-
Threonyl-tRNA synthetase
Q9YFY3
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
Escherichia coli MG1655
-
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
Threonyl-tRNA synthetase
-
-
ThrRS
-
-
-
-
ThrRS
Q9YDW0
-
ThrRS
Escherichia coli MG1655
-
-
-
TRS
-
-
-
-
TRS
Q9YFY3
-
mitochondrial threonyl-tRNA synthetase
-
-
additional information
-
the enzyme belongs to the class II aminoacyl-tRNA synthetases
CAS REGISTRY NUMBER
COMMENTARY
9023-46-5
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
isozyme ThrRS-1, K1, extreme thermophilic archaeon
GenBank
Manually annotated by BRENDA team
isozyme ThrRS-2, K1, extreme thermophilic archaeon
GenBank
Manually annotated by BRENDA team
several creanarchaea including Aeropyrum pernix K1 and Sulfolobus tokodaii strain 7 contain two genes encoding either the catalytic or the editing domain of ThrRS
-
-
Manually annotated by BRENDA team
parental GAT-cell line and borrelidin-resistant derivative 2000A
-
-
Manually annotated by BRENDA team
class II enzyme
-
-
Manually annotated by BRENDA team
class II enzyme, recombinantly overexpressed and purified enzyme
-
-
Manually annotated by BRENDA team
K12, wild-type and borrelidin-resistant mutant
-
-
Manually annotated by BRENDA team
overproducing strain
-
-
Manually annotated by BRENDA team
several strains, overview, gene thrS
-
-
Manually annotated by BRENDA team
Escherichia coli MG1655
-
-
-
Manually annotated by BRENDA team
Escherichia coli overproducing
overproducing strain
-
-
Manually annotated by BRENDA team
extreme halophilic archaeon
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae D273-10B
D273-10B
-
-
Manually annotated by BRENDA team
several creanarchaea including Aeropyrum pernix K1 and Sulfolobus tokodaii strain 7 contain two genes encoding either the catalytic or the editing domain of ThrRS
-
-
Manually annotated by BRENDA team
overproduced in Escherichia coli
-
-
Manually annotated by BRENDA team
strain HB8
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
-
tRNAThr phylogenetic analysis, overview
evolution
-
evolutionary development of unusual tRNAThr1, containing an enlarged 8-nt anticodon loop, which is required to reassign CUN codons from leucine to threonine, tRNAThr phylogenetic analysis, overview. MST1 has co-evolved with tRNAThr, which evoled from tRNAHis
malfunction
-
reactive oxygen species cause editing defect and misacylation by WT ThrRS. H2O2-induced Ser-tRNAThr formation causes protein mistranslation
malfunction
Escherichia coli MG1655
-
reactive oxygen species cause editing defect and misacylation by WT ThrRS. H2O2-induced Ser-tRNAThr formation causes protein mistranslation
-
metabolism
-
in certain yeast mitochondria, CUN codons are reassigned from leucine to threonine, which requires an unusual tRNAThr with an enlarged 8-nt anticodon loop, tRNAThr1. But in Candida albicans, the CUN codons remain assigned to leucine, and MST1 cannot threonylate tRNAThr
metabolism
-
in certain yeast mitochondria, CUN codons are reassigned from leucine to threonine, which requires an unusual tRNAThr with an enlarged 8-nt anticodon loop, tRNAThr1
physiological function
Q9YDW0, Q9YFY3
ApThrRS-1 catalyses only the aminoacylation of the cognate tRNA; ApThrRS-2 is necessary for trans-editing, like the deaminoacylation of a misacylated serine moiety at the CCA terminus
physiological function
-
the final localisation of Thr-tRNA synthetase, expressed as a precursor protein, depends on the dual targeting peptide ThrRS-dTP
physiological function
-
amino acid discrimination does not occur at the aminoacyl transfer step. pre-Transfer hydrolysis contributes to proofreading only when the rate of transfer is slowed significantly. Thus, the relative contributions of pre- and posttransfer editing in ThrRS are subject to modulation by the rate of aminoacyl transfer
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-Chloroadenosine 5'-triphosphate + L-threonine + tRNAThr
?
show the reaction diagram
-
-
-
-
-
2-Chloroadenosine 5'-triphosphate + L-threonine + tRNAThr
?
show the reaction diagram
-
-
-
-
-
ATP + 3-hydroxynorvaline + tRNAThr
AMP + diphosphate + 3-hydroxynorvalyl-tRNAThr
show the reaction diagram
-
the specificity constant kcat/KM for beta-hydroxynorvaline is only 20-30fold less than that of cognate threonine, amino acid activation is the potential rate-limiting step of b3-hydroxynorvaline aminoacylation
-
-
?
ATP + hydroxynorvaline + tRNAThr
AMP + diphosphate + hydroxynorvalyl-tRNAThr
show the reaction diagram
P0A8M3
10-70% less active than with L-threonine
-
?
ATP + L-isoleucine + tRNAIle
AMP + diphosphate + L-isoleucyl-tRNAIle
show the reaction diagram
-
-, the reaction catalyzed by the enzyme plays an important role in the transport of aminoacylated tRNAs from the nucleus to the cytoplasm
-
?
ATP + L-serine + tRNAThr
AMP + diphosphate + L-seryl-tRNAThr
show the reaction diagram
Q980D1, -
-
-
-
-
ATP + L-serine + tRNAThr
AMP + diphosphate + L-seryl-tRNAThr
show the reaction diagram
-
low activity
-
-
r
ATP + L-serine + tRNAThr
AMP + diphosphate + L-seryl-tRNAThr
show the reaction diagram
P0A8M3
1000fold less active than with L-threonine
-
?
ATP + L-serine + tRNAThr
AMP + diphosphate + L-seryl-tRNAThr
show the reaction diagram
-
very low activity with the wild-type enzyme
-
?
ATP + L-serine + tRNAThr
AMP + diphosphate + L-seryl-tRNAThr
show the reaction diagram
Q58597
L-serine is a poor substrate for the wild-type enzyme
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q8NW68, -
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
P0A8M3
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-, Q9UZ14
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q9YDW0, Q9YFY3
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q9YDW0, Q9YFY3
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q980D1, -
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q58597
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q9YDW0, Q9YFY3
secondary structure of the tRNAThr, enzyme utilizes substrates from archaeon with discriminator base U73 and Escherichia coli with discriminator base A73
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
secondary structure of the tRNAThr, specific for archaeal tRNAThr with discriminator base U73, no activity with one of Escherichia coli with discriminator base A73
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q8NW68, -
substrate binding induces conformational changes
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
tRNA aminoacylation
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
regulatory mechanism
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
the reaction catalyzed by the enzyme plays an important role in the transport of aminoacylated tRNAs from the nucleus to the cytoplasm
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
determination of substrate binding sites, catalytic sites, and catalytic mechanism
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
editing mechanism
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
via AMP-L-threonine-enzyme intermediate in a two-step process
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
the enzyme acts as both an enzyme and a regulator of gene expression, it aminoacylates tRNAThr isoacceptors and binds to its own mRNA, inhibiting its translation, overview
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-, Q9UZ14
the main chains atoms of Tyr119 and Tyr120 are sufficient to prevent the deacylation of Thr-tRNAThr
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
the zinc atom in the active site is essential for the recognition of threonine
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
for cognate threonine, amino acid activation is likely to be the rate-limiting step. The inability of wild-type ThrRS to prevent utilization of beta-hydroxynorvaline as a substrate illustrates that the naturally occurring enzyme lacks the capability to effectively discriminate against nonproteogenic amino acids that are not encountered under normal physiological conditions
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
MST1 can attach threonine to both tRNAThr1 and the regular tRNAThr2, but not to the wild-type tRNAHis. But a loss of the first nucleotide G-1 in tRNAHis converts it to a substrate for MST1
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Escherichia coli overproducing
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Saccharomyces cerevisiae D273-10B
-
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Escherichia coli MG1655
-
-
-
-
?
ATP + L-threonine + tRNAThr
?
show the reaction diagram
-
catalyzes the attachment of threonine onto its cognate tRNA molecule, prior to participation of the aminoacylated tRNA in the protein synthesis
-
-
-
Formycin 5'-triphosphate + L-threonine + tRNAThr
?
show the reaction diagram
-
-
-
-
-
Formycin 5'-triphosphate + L-threonine + tRNAThr
?
show the reaction diagram
-
-
-
-
-
additional information
?
-
-
enzyme can interact with high-MW RNAs
-
-
-
additional information
?
-
-
the enzyme examines side chain structures of amino acids in 4 recognition steps. For each step the enzyme uses special distinct structures or conformations of the binding cleft, specificity with regard to amino acids, discrimination factors
-
-
-
additional information
?
-
-
enzyme also has a regulatory function by binding the so-called operator site located in the leader of its own mRNA and thereby inhibits translational initiation by competing with ribosome binding
-
?
additional information
?
-
P0A8M3
no activity with L-valine, determination of amino acid activation and discriminating editing mechanism
-
?
additional information
?
-
-
the enzyme represses the translation of its own mRNA by binding to an operator located upstream of the initiation codon thereby using the recognition mode of the tRNA anticodon loop to initiate binding
-
?
additional information
?
-
-
aminoacyl-tRNA is channeled in vivo by probably direct transfer to elongation factor I
-
?
additional information
?
-
-
regulation mechanism, 2 essential steps of regulation are operator recognition and inhibition of ribosome binding performed by different domains of the enzyme
-
?
additional information
?
-
Q980D1, -
a freestanding proofreading domain is required for protein synthesis quality control in archaea
-
-
-
additional information
?
-
Q58597
a freestanding proofreading domain is required for protein synthesis quality control in archaea
-
-
-
additional information
?
-
-
structure-based evolutionary considerations
-
-
-
additional information
?
-
-
the enzyme needs to discriminate between threonine, serine, and valine in vivo, mechanism of proofreading of threonyl-tRNA synthetase at atomic resolution, overview
-
-
-
additional information
?
-
Q980D1, -
L-serine is a poor substrate for the wild-type enzyme, the N-terminal enzyme domain is responsible for editing
-
-
-
additional information
?
-
Q58597
the N-terminal enzyme domain is responsible for editing
-
-
-
additional information
?
-
-, Q9UZ14
mischarging of the enzyme with noncognate amino acids, overview, post-transfer editing mechanism of the D-aminoacyl-tRNA deacylase-like domain in the archaeal threonyl-tRNA synthetase, mechanistic insights into the removal of noncognate L-serine from tRNAThr, M129 is responsible for enantiomeric selection in DTD, Glu134 is involved in fixing the seryl moiety in the active site, overview
-
-
-
additional information
?
-
-
addition of first nucleotide G-1 to tRNAThr1 allows efficient histidylation by histidyl-tRNA synthetase
-
-
-
additional information
?
-
-
in the pre-steady state, asymmetric activation of cognate threonine and noncognate serine is observed in the active sites of dimeric ThrRS, with similar rates of activation. In the absence of tRNA, seryl-adenylate is hydrolyzed 29old faster by the ThrRS catalytic domain than threonyl-adenylate. The rate of seryl transfer to cognate tRNA is only 2fold slower than threonine
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-isoleucine + tRNAIle
AMP + diphosphate + L-isoleucyl-tRNAIle
show the reaction diagram
-
the reaction catalyzed by the enzyme plays an important role in the transport of aminoacylated tRNAs from the nucleus to the cytoplasm
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q8NW68, -
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
P0A8M3
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-, Q9UZ14
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q9YDW0, Q9YFY3
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q9YDW0, Q9YFY3
-
-
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q980D1, -
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Q58597
-
-
-
r
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
regulatory mechanism
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
the reaction catalyzed by the enzyme plays an important role in the transport of aminoacylated tRNAs from the nucleus to the cytoplasm
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
-
the enzyme acts as both an enzyme and a regulator of gene expression, it aminoacylates tRNAThr isoacceptors and binds to its own mRNA, inhibiting its translation, overview
-
-
?
ATP + L-threonine + tRNAThr
?
show the reaction diagram
-
catalyzes the attachment of threonine onto its cognate tRNA molecule, prior to participation of the aminoacylated tRNA in the protein synthesis
-
-
-
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
Escherichia coli MG1655
-
-
-
-
?
additional information
?
-
-
aminoacyl-tRNA is channeled in vivo by probably direct transfer to elongation factor I
-
?
additional information
?
-
-
regulation mechanism, 2 essential steps of regulation are operator recognition and inhibition of ribosome binding performed by different domains of the enzyme
-
?
additional information
?
-
Q980D1, -
a freestanding proofreading domain is required for protein synthesis quality control in archaea
-
-
-
additional information
?
-
Q58597
a freestanding proofreading domain is required for protein synthesis quality control in archaea
-
-
-
additional information
?
-
-
structure-based evolutionary considerations
-
-
-
additional information
?
-
-
the enzyme needs to discriminate between threonine, serine, and valine in vivo, mechanism of proofreading of threonyl-tRNA synthetase at atomic resolution, overview
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
K+
-
activates
K+
-
optimal concentration: 150 mM KCl
Mg2+
-
Mg2+ can be substituted by Ca2+ and Mn2+. Zn2+ and Ni2+ can only very poorly replace Mg2+; required
Mg2+
-
Mg2+ can be substituted by Ba2+, Ca2+, Mn2+ and Co2+; required
Mg2+
-
optimal concentration: 20 mM MgCl2
Mg2+
Q8NW68, -
required
Mg2+
-
binding and structure modeling, ligand coordination
NH4+
-
partial activation
Phosphorus
-
phosphoprotein containing phosphoserine, activity can be modulated by reversible phosphorylation
Rb+
-
partial activation
Zn2+
-
enzyme contains 1 Zn2+ per subunit
Zn2+
-
used to discriminate against the isosteric valine at the activation step
Zn2+
-
essential, binds at the active site
Zn2+
-
mediates amino acid discrimination of the enzyme, located in the active site, formation of a pentacoordinate intermediatewith both the amino group and the side chain hydroxyl of L-threonine
Zn2+
Q9YDW0, Q9YFY3
mediates amino acid discrimination, binding model
Zn2+
-
assures that no valine is bound
Zn2+
-
binding and structure modeling, ligand coordination
Zn2+
-
zinc binding in the catalytic site, the active site zinc atom is essential for the recognition of threonine, three amino acids forming the zinc-binding site, overview
Zn2+
Q9YDW0, Q9YFY3
;
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2'-Deoxyadenosine 5'-triphosphate
-
-
2'-O-Methyladenosine 5'-triphosphate
-
-
3'-Deoxyadenosine 5'-triphosphate
-
-
3'-O-Methyladenosine 5'-triphosphate
-
-
Borrelidin
-
-
Borrelidin
-
inhibition mechanism via conformational change abolishing the activation of threonine, a unique hydrophobic cluster near the active site contributes to differences in borrelidin inhibition among threonyl-tRNA synthetases of different origin, comparison, overview
Borrelidin
-
slowly but tight binding, noncompetitive with respect to threonine and ATP, inhibition mechanism via conformational change abolishing the activation of threonine, a unique hydrophobic cluster near the active site contributes to differences in borrelidin inhibition among threonyl-tRNA synthetases of different origin, comparison, overview
Borrelidin
-
inhibition mechanism via conformational change abolishing the activation of threonine, a unique hydrophobic cluster near the active site contributes to differences in borrelidin inhibition among threonyl-tRNA synthetases of different origin, comparison, overview
operator mRNA domain 2
-
-
-
Purineriboside 5'-triphosphate
-
-
Tubercidin 5'-triphosphate
-
-
Zn2+
-
inhibits the editing reaction
hydrogen peroxide
-
oxidizes cysteine182 residue critical for editing, which leads to Ser-tRNAThr formation and protein mistranslation that impaired growth of Escherichia coli. Presence of major heat shock proteases is required to allow cell growth in medium containing serine and hydrogen peroxide, which suggests that the mistranslated proteins are misfolded
additional information
-
reactive oxygen species cause editing defect and misacylation by WT ThrRS
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
spermine
-
has a stimulatory and synergistic effect with Mg2+, cannot replace Mg2+ in activation
spermine
-
no effect on the overall reaction as well as on the rate of ATP-diphosphate exchange. At low Mg2+ (1 mM) concentration spermine activates 8fold ATP-diphosphate exchange and 23fold the threonyl-tRNA synthesis
additional information
-
3-hydroxynorvaline enhances the ATPase function of the synthetic site, at a rate not increased by nonaminoacylatable tRNA
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.4
-
2-Chloroadenosine 5'-triphosphate
-
-
0.4
-
2-Chloroadenosine 5'-triphosphate
-
ATP
0.267
-
ATP
-
wild-type enzyme, pH 7.2, 37C
0.387
-
ATP
-
mutant enzyme W434Y, pH 7.2, 37C
0.5
-
Formycin 5'-triphosphate
-
-
0.5
-
Formycin 5'-triphosphate
-
threonine
1.95
-
hydroxynorvaline
-
pH 7.2, 37C, wild-type enzyme
7
-
hydroxynorvaline
-
pH 7.2, 37C, truncated enzyme DELTAN
25
-
L-serine
Q58597
pH 7.2, 60C, recombinant wild-type enzyme
55
-
L-serine
Q980D1, -
pH 7.2, 60C, recombinant wild-type enzyme
81.5
-
L-serine
-
pH 7.2, 37C, wild-type enzyme
142
-
L-serine
-
pH 7.2, 37C, truncated enzyme DELTAN
0.00003
-
L-threonine
-
wild-type enzyme complementing the null mutant
0.09
-
L-threonine
-
pH 7.5, 65C, wild-type enzyme
0.1
-
L-threonine
-
pH 7.5, 65C, wild-type enzyme
0.1
-
L-threonine
-
pH 7.5, 55C, wild-type enzyme
0.1
-
L-threonine
Q58597
pH 7.2, 60C, recombinant wild-type enzyme
0.11
-
L-threonine
-
pH 7.2, 37C, wild-type enzyme
0.11
-
L-threonine
-
pH 7.5, 37C, wild-type enzyme
0.11
-
L-threonine
Q980D1, -
pH 7.2, 60C, recombinant wild-type enzyme
0.18
-
L-threonine
-
pH 7.2, 37C, truncated enzyme DELTAN
0.201
-
L-threonine
-
wild-type enzyme, pH 7.2, 37C
0.00189
-
threonine
-
-
0.00003
-
tRNAThr
-
wild-type enzyme
0.000037
0.00007
tRNAThr
-
of Thermus thermophilus, , depending on temperature
0.00005
-
tRNAThr
-
of E. coli
0.000085
-
tRNAThr
-
-
0.00014
-
tRNAThr
-
of E. coli
0.00029
-
tRNAThr
-
pH 7.2, 37C, tRNAThr1, MST1
0.00043
-
tRNAThr
-
truncated enzyme core DELTAN
0.00044
-
tRNAThr
-
pH 7.2, 37C, tRNAThr2, MST1
0.0015
-
tRNAThr
-
pH 7.2, 37C, tRNAThr1(G-1), MST1
0.0015
-
tRNAThr
-
pH 7.2, 37C, ThrRS, aminoacylation with Thr
0.012
-
tRNAThr
-
-
0.179
-
tRNAThr
-
of yeast
0.897
-
L-threonine
-
mutant enzyme W434Y, pH 7.2, 37C
additional information
-
additional information
-
-
-
additional information
-
additional information
-
Km values of variants of tRNAThr transcripts
-
additional information
-
additional information
-
kinetics and kinetic mechanism
-
additional information
-
additional information
-
mutant enzymes complementing the null mutant
-
additional information
-
additional information
-
steady-state kinetic measurement
-
additional information
-
additional information
-
presteady-state and steady-state kinetic measurement
-
additional information
-
additional information
-
steady-state kinetic measurement
-
additional information
-
additional information
Q58597
kinetics of recombinant wild-type and mutant enzymes with threonine and serine
-
additional information
-
additional information
Q980D1, -
kinetics of recombinant wild-type and mutant enzymes with threonine and serine
-
additional information
-
additional information
-
pre-steady-state kinetics, kinetics of ATPase activity in presence of 3-hydroxynorvaline, overview
-
additional information
-
additional information
-
kinetics of MST1 with different tRNAs, overview
-
additional information
-
additional information
-
kinetics of diphosphate exchange activities and threonylation of tRNAThr of ThrRS with or without H2O2 treatment, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
42
-
ATP
-
mutant enzyme W434Y, pH 7.2, 37C
90
-
ATP
-
wild-type enzyme, pH 7.2, 37C
21
-
hydroxynorvaline
-
pH 7.2, 37C, truncated enzyme DELTAN
22
-
hydroxynorvaline
-
pH 7.2, 37C, wild-type enzyme
1.3
-
L-serine
Q58597
pH 7.2, 60C, recombinant wild-type enzyme
12.3
-
L-serine
Q980D1, -
pH 7.2, 60C, recombinant wild-type enzyme
26
-
L-serine
-
pH 7.2, 37C, wild-type enzyme
30
-
L-serine
-
pH 7.2, 37C, truncated enzyme DELTAN
0.64
-
L-threonine
-
wild-type enzyme complementing the null mutant
1.2
-
L-threonine
-
pH 7.5, 65C, wild-type enzyme
1.9
-
L-threonine
Q58597
pH 7.2, 60C, recombinant wild-type enzyme
3
6
L-threonine
-
pH 7.2, 37C, wild-type enzyme
3.8
-
L-threonine
-
pH 7.5, 65C, wild-type enzyme
6.08
-
L-threonine
-
wild-type enzyme complementing the null mutant
13.5
-
L-threonine
Q980D1, -
pH 7.2, 60C, recombinant wild-type enzyme
16
-
L-threonine
-
pH 7.5, 55C, wild-type enzyme
30
-
L-threonine
-
mutant enzyme W434Y, pH 7.2, 37C
33
-
L-threonine
-
pH 7.5, 37C, wild-type enzyme
37
-
L-threonine
-
pH 7.2, 37C, truncated enzyme DELTAN
0.53
-
RNAThr
-
tRNAThr of E. coli
-
1.73
-
threonyl-tRNA
-
-
-
0.00157
-
tRNAThr
-
pH 7.2, 37C, tRNAThr1(G-1), MST1
0.0383
-
tRNAThr
-
pH 7.2, 37C, tRNAThr2, MST1
0.0467
-
tRNAThr
-
pH 7.2, 37C, tRNAThr1, MST1
0.05
-
tRNAThr
-
pH 7.2, 37C, ThrRS, aminoacylation with Thr
0.21
-
tRNAThr
-
tRNAThr of E. coli
0.52
0.7
tRNAThr
-
tRNAThr of Thermus thermophilus
0.037
0.23
tRNAThr of Thermus thermophilus
-
-
-
90
-
L-threonine
-
wild-type enzyme, pH 7.2, 37C
additional information
-
additional information
-
-
-
additional information
-
additional information
-
mutant enzymes complementing the null mutant
-
additional information
-
additional information
-
rate constants for adenylate synthesis by ThrRS in the absence of tRNA, overview
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.093
-
tRNAThr
-
pH 7.2, 37C, tRNAThr1(G-1), MST1
17520
38.3
-
tRNAThr
-
pH 7.2, 37C, ThrRS, aminoacylation with Thr
17520
90
-
tRNAThr
-
pH 7.2, 37C, tRNAThr2, MST1
17520
171.7
-
tRNAThr
-
pH 7.2, 37C, tRNAThr1, MST1
17520
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.000004
-
Borrelidin
-
pH 7.5, 37C, wild-type enzyme
0.0000045
-
Borrelidin
-
pH 7.5, 55C, wild-type enzyme
0.006
-
Borrelidin
-
above, pH 7.5, 65C, wild-type enzyme
0.006
-
Borrelidin
-
pH 7.5, 37C, mutant L489W
0.006
-
Borrelidin
-
above, pH 7.5, 65C, wild-type enzyme
additional information
-
additional information
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.2
-
-
assay at
7.2
-
Q58597
assay at
7.2
-
Q980D1, -
assay at
7.2
-
-
asay at
7.2
-
-
assay at
7.4
-
-
presteady-state, assay at
7.5
-
Q9YDW0, Q9YFY3
assay at
7.5
-
-
assay at
7.5
-
-
assay at
7.5
-
-
steady-state, assay at
7.5
-
-
assay at
7.7
-
Q8NW68, -
assay at
7.7
-
-
aminoacylation assay at
8
-
-
assay at
8.4
-
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-
presteady-state, assay at
37
-
-
assay at
37
-
Q9YDW0, Q9YFY3
assay at
37
-
-
steady-state, assay at
37
-
-
assay at
37
-
-
aminoacylation assay at
37
-
-
assay at
37
-
-
assay at
37
-
-
assay at
50
-
-
assay at
60
-
Q58597
assay at
60
-
Q980D1, -
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
cell line LCC-RK1
Manually annotated by BRENDA team
-
cell line CRL-1781
Manually annotated by BRENDA team
-
parental GAT- and borrelidin-resistant 2000A
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
major part, enzyme participates in a multienzyme complex, not as large and stable as the one from nucleus
Manually annotated by BRENDA team
Saccharomyces cerevisiae D273-10B
-
-
-
Manually annotated by BRENDA team
-
0.4% of total activity in the cell, enzyme participates in a large and stable multienzyme complex
Manually annotated by BRENDA team
-
0.15% of total activity in the cell, enzyme participates in a large and stable multienzyme complex
Manually annotated by BRENDA team
-
association of the enzyme with high MW cellular component
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Pyrococcus abyssi (strain GE5 / Orsay)
Pyrococcus abyssi (strain GE5 / Orsay)
Pyrococcus abyssi (strain GE5 / Orsay)
Pyrococcus abyssi (strain GE5 / Orsay)
Pyrococcus abyssi (strain GE5 / Orsay)
Pyrococcus abyssi (strain GE5 / Orsay)
Pyrococcus abyssi (strain GE5 / Orsay)
Pyrococcus abyssi (strain GE5 / Orsay)
Pyrococcus abyssi (strain GE5 / Orsay)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6600
-
-
ThrRS-dual targeting peptide, determined by SDS-PAGE
30860
-
-
predicted from cDNA
45000
-
-
predicted from cDNA
53120
-
-
predicted from cDNA
63110
-
-
predicted from cDNA
150000
-
-
-
150000
-
-
gel filtration
150000
-
-
gel filtration
150000
-
-
gel filtration
154000
-
-
sedimentation equilibrium measurement
164000
-
-
PAGE at different gel concentrations
170000
-
-
gel filtration
195000
-
-
mitochondrial, gel filtration
206000
-
-
cytoplasmic, gel filtration
additional information
-
-
nucleotide and deduced amino acid sequence
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
2 * 77000, SDS-PAGE under denaturing conditions
dimer
-
2 * 76000, SDS-PAGE
dimer
-
2 * 82000, SDS-PAGE
dimer
-
2 * 85000, SDS-PAGE
dimer
-
2 * 86000, SDS-PAGE
dimer
Escherichia coli overproducing
-
2 * 76000, SDS-PAGE
-
homodimer
-
ApThrRS-1 forms a homodimer of subunits which are related to each other by crystallographic twofold rotation symmetry along the a axis. The amino-acid residues at the interface interact with each other through hydrogen bonds and van der Waals contacts between the two subunits. This structural feature is consistent with the dimer formation of the class II ThrRSscrystal structure; ApThrRS-2 might form a dimer between the editing domains, as the catalytic domain for dimerization is missing. This is consistent with the results of gel-filtration experiment, which shows that the molecular size of ApThrRS-2 is equivalent to twice that of the subunit
homodimer
Q9YDW0, Q9YFY3
-
additional information
Q9YDW0, Q9YFY3
structure schematic model
additional information
Q58597
the N-terminal enzyme domain is responsible for editing
additional information
Q980D1, -
the N-terminal enzyme domain is responsible for editing
additional information
-
three-dimensional structure modeling of the N-terminal domain by fold recognition and sequence analysis, determination of substrate binding site and catalytic site
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystallized by the hanging-drop vapour diffusion method; crystallized by the hanging-drop vapour diffusion method. Diffraction data are collected and the structure of a selenomethionine-labelled Aeropyrum pernix type-1 ThrRS crystal is solved using the multiple anomalous dispersion method
-
the structure of ApThrRS-1 is determined at 2.3 A resolution
Q9YDW0, Q9YFY3
crystal structure, PDB code 1EVL, at 1.55 A resolution used for reaction mechanism analysis and molecular modeling
-
enzyme core DELTAN complexed with the essential operator domain mRNA, precipitation with 1.9 M ammonum sulfate in sodium cacodylate, pH 6.6, 4C from solution containing 0.088 mM enzyme mutant DELTAN, 0.2 mM operator mRNA domain 2, 10 mM MgCl2, 10 mM substrate analogue 5'-O-(N-(L-threonyl)-sulfamoyl)adenosine, crystals are soaked in 25% v/v glycerol, X-ray diffraction structure determination at 3.6 A resolution and structure analysis
-
enzyme in complex with tRNAThr, hanging drop method, pH 6.5, solution containing ammonium acetate, ATP, and PEG 4000 as precipitant, X-ray diffraction structure determination at 2.9 A resolution and analysis
-
lamdaN-threonine-tRNA ligase complexed with Ser-AMS, X-ray diffraction structure determination at 1.65 A resolution and analysis
-
purified recombinant N-terminal part of the enzyme, i.e. N1 and N2 domains comprising residues 1-65 and 66-225 in one fragment, X-ray diffraction structure determination and analysis at 1.5 A resolution, structure modeling
-
X-ray structure determination at 2.9 A resolution, structure analysis of the wild-type enzyme and truncated mutant lamdaN in complex with L-threonine and L-serine
-
purified recombinant D-aminoacyl-tRNA deacylase-like domain with bound Ser3AA, hanging drop vapor diffusion method, mixing of equal volumes of protein, ligand and reservoir solution, crystallization from 0.2 M (NH4)2SO4, 0.1 M sodium cacodylate, pH 6.5, and 30% PEG 8000 and 0.1 M HEPES, pH 7.0, and 25% PEG 3350, purified recombinant D-aminoacyl-tRNA deacylase-like domain with bound SerAMS, using 0.1 M Bis-Tris, pH 6.5, and 25% PEG 3350, and the serine Pab-NTDL-serine cocrystals from 0.1 M HEPES pH 7.0 and 25% PEG 8000, X-ray diffraction structure determination and analysis at 1.86-2.25 A resolution, overview
-
purified ultrafiltrated recombinant residues 1-183, hanging drop vapour diffusion method, protein solution, containing 50 mM Tris-HCl, pH 7.5, and 50 mM NaCl, drops of varying volumes against 0.75 ml reservoir solution, at room temperature or at 4C, preliminary X-ray diffraction for structure determination at 1.95 A resolution
-
enzyme complexed with ATP and threonine and complexed with substrate analogue 5'-O-(N-(L-threonyl)-sulfamoyl)adenosine, hanging drop method, 15 mg/ml protein in 20 mM HEPES, pH 7.2, 2 mM MgCl2, 100 mM KCl, in a 1:1:1 mixture with ligand solution containing 10 mM MgCl2, 10 mM ATP or 1 mM Thr-AMS, and well solution containing 50 mM Tris-HCl, pH 7.5-8.5, PEG 8000 12-14% w/v, and ammonium acetate or potassium chloride in the range 0.2-0.4 M, 2-3 days at 4C, X-ray diffraction structure determination at resolution 2.8-3.2 A, structure analysis, modeling
Q8NW68, -
crystallized by the hanging-drop vapour diffusion method
-
enzyme overproduced in Escherichia coli
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
60
-
-
12 min, incubation without substrates, 50% inactivation
additional information
-
-
threonine does not influence the rate of heat inactivation. ATP and tRNA labilize
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80C, stable for more than 18 months
-
-80C, about 30% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
;
Q9YDW0, Q9YFY3
using a RESOURCE S column
-
recombinant His-tagged enzyme from Escherichia coli mutant by nickel affinity chromatography
-
recombinant His-tagged enzyme from strain IBPC6881, to homogeneity
-
recombinant His-tagged ThrRS by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzyme from Escherichia coli mutant by nickel affinity chromatography
-
recombinant non-tagged N1 and N2 domains comprising residues 1-65 and 66-225 in one fragment by ion exchange chromatography, ammonium sulfate fractionation, hydrophobic interaction chromatography, and ultrafiltration
-
recombinant His-tagged mutant H385A
-
recombinant His-tagged enzyme from Escherichia coli mutant by nickel affinity chromatography
-
recombinant His-tagged enzyme from Escherichia coli mutant by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant D-aminoacyl-tRNA deacylase-like domains from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
-
recombinant residues 1-183 from Escherichia coli strain BL21(DE3) by ion exchange chromatography, ammonium sulfate fractionation and gel filtration to homogeneity
-
5SrRNA-L5 protein complex containing both ThrRS and HisRS
-
recombinant from Escherichia coli
Q8NW68, -
recombinant His-tagged enzyme from Escherichia coli mutant by nickel affinity chromatography
-
overproduced in Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
DNA sequence analysis
Q9YDW0, Q9YFY3
for expression in Escherichia coli cells; for expression in Escherichia coli cells
Q9YDW0, Q9YFY3
the two types of ThrRS from the crenarchaea Aeropyrum pernix and Sulfolobus tokodaii are overexpressed in Eschericha coli
-
deletion constructs of the ThrRS-dual targeting peptide, ThrRS-dTP, are generated, consisting of amino acid residues 1-50, 1-60, or 24-68, ligated into the pCR Blunt II TOPO vector as a fusion protein with GFP located C-terminally, for a GST-ThrRS-dTP(1-60) construct the vector pGEX-6P-2 is used
-
expression of His-tagged enzyme in an enzyme-deficient Escherichia coli mutant, complementation analysis
-
expression of His-tagged ThrRS
-
expression of His-tagged wild-type and mutant enzyme in an enzyme-deficient Escherichia coli mutant, complementation analysis
-
expression of non-tagged N1 and N2 domains comprising residues 1-65 and 66-225 in one fragment in strain BL21
-
overexpression in strain IBPC6881 as His-tagged enzyme
-
overexpression of wild-type and mutants in the null mutant strain
-
overexpression of His-tagged mutant enzymes
-
expression of His-tagged enzyme in an enzyme-deficient Escherichia coli mutant, complementation analysis
-
expression of His-tagged enzyme in an enzyme-deficient Escherichia coli mutant, complementation analysis
-
gene thrS, expression of wild-type and mutant enzymes in Escherichia coli strain BL21
Q58597
DNA and amino avid sequence analysis
-
DNA and amino acid sequence determination of residues 1-183, expression in Escherichia coli strain BL21(DE3)
-
overexpression of His-tagged wild-type and mutant D-aminoacyl-tRNA deacylase-like domains in Escherichia coli strain BL21(DE3), mutant M129K is located in inclusion bodies
-
overexpression in Escherichia coli
Q8NW68, -
expression of His-tagged enzyme in an enzyme-deficient Escherichia coli mutant, complementation analysis
-
gene thrS, expression of wild-type and mutant enzymes in Escherichia coli strain BL21
Q980D1, -
the two types of ThrRS from the crenarchaea Aeropyrum pernix and Sulfolobus tokodaii are overexpressed in Eschericha coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D180A
-
charging of tRNAThr with serine, mutant is no longer able to rapidly deacetylate Ser-tRNAThr
D435A
-
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
D549A
-
modified interation of anticodon loop/C-ter domain, activity similar to the wild-type
E258K
-
modified interation with the superrepressor, no activity
E259K
-
modified interation with the superrepressor, unaltered activity
E600A
-
modified interation of anticodon loop/C-ter domain, 710fold increased activity
H309A
-
site-directed mutagenesis, mutant shows highly increased Ki for inhibitor borrelidin compared to the wild-type enzyme
H337A
-
site-directed mutagenesis, mutant shows increased Ki for inhibitor borrelidin compared to the wild-type enzyme
H73A
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme, and a 2fold higher rate of ATP consumption relative to the rate of Ser-tRNAThr synthesis
H73A/H309A
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme, and a 2fold higher rate of ATP consumption relative to the rate of Ser-tRNAThr synthesis
H73A/H77A
-
charging of tRNAThr with serine, mutant is no longer able to deacetylate Ser-tRNAThr
K246A
-
modified interation of acceptor stem and catalytic domain, 2.9fold increased activity
K249A
-
modified interation of acceptor stem and catalytic domain, 3.5fold increased activity
K577A
-
modified interation of anticodon loop/C-ter domain, 118fold increased activity
N324A
-
modified interation of cross-subunit contacts, 3.5fold increased activity
N502A
-
modified interation of cross-subunit contacts, 2.1fold increased activity
N575A
-
modified interation of anticodon loop/C-ter domain, 9.4fold increased activity
P296A
-
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
P296S
-
site-directed mutagenesis, mutant shows slightly increased Ki for inhibitor borrelidin compared to the wild-type enzyme
P335A
-
site-directed mutagenesis, mutant shows increased Ki for inhibitor borrelidin compared to the wild-type enzyme
P464A
-
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
R282A
-
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
R349A
-
modified interation of cross-subunit contacts, 42fold increased activity
R583H
-
modified interation of anticodon loop/C-ter domain, no activity
R609A
-
modified interation of anticodon loop/C-ter domain, 35fold activity
S347A
-
modified interation of cross-subunit contacts, similar to the wild-type
S367A
-
modified interation of acceptor stem and catalytic domain, 11fold increased activity
S429A
-
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
T307A
-
site-directed mutagenesis, mutant shows increased Ki for inhibitor borrelidin compared to the wild-type enzyme
W434Y
-
reduced activity, Trp434 is involved in conformational changes during substrate binding
Y205F
-
modified interation of acceptor stem and N-terminal domain, 7.7fold increased activity
Y219F
-
modified interation of acceptor stem and N-terminal domain, similar to the wild-type
Y313A
-
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
Y348F
-
modified interation of cross-subunit contacts, 6.5fold increased activity
H385A
-
site-directed mutagenesis, the mutant shows altered substrate specificity, overview
H385N
-
site-directed mutagenesis, the mutant shows altered substrate specificity, overview
E134A
-
site-directed mutagenesis, the mutation has no effect on the deacylation activity
H83A
-
site-directed mutagenesis, the mutant possesses the editing activity albeit with a slower rate compared to the wild-type enzyme
K121A
-
no expression for the alanine mutant
K121M
-
site-directed mutagenesis, substitution of Lys121 to serine does not abolish Ser-tRNAThr deacylation activity
K121S
-
site-directed mutagenesis, substitution of Lys121 to serine results in a complete abolition of Ser-tRNAThr deacylation activity
M129K
-
site-directed mutagenesis, the mutant shows binding not only to L-serine but also to a variety of other L-amino acids that are tested in addition to binding to various D-amino acids, overview
additional information
-
borrelidin-resistant derivative cell line 2000A
L489W
-
site-directed mutagenesis, mutant has a reduced space of the hydrophobic cluster near the active site resulting in a 1500fold increase in Ki for inhibitor borrelidin compared to the wild-type enzyme
additional information
-
borrelidin-resistant mutant
additional information
-
construction of chromosomal disruption null mutant strain with no activity, construction of a truncated mutant lacking the N1 and N2 domains, 93.5fold increased activity
additional information
-
truncated lamdaN-enzyme mutant, lacking the N-terminal domains N1 and N2, produces Ser-tRNAThr, reduced activity and altered substrate recognition compared to the wild-type which does nearly not incorporate serine
additional information
-
construction of a truncated enzyme: core domain DELTAN comprising residues 242-642
H385Y
-
site-directed mutagenesis, the mutant shows altered substrate specificity, overview
additional information
-
mutations of any of the three amino acids forming the zinc-binding site inactivate the enzyme and have a dominant negative effect on growth if the corresponding genes are placed on a multicopy plasmid, not due to the formation of inactive heterodimers, the titration of tRNAThr by an inactive enzyme, or its misaminoacylation but is, rather, due to the regulatory function of threonyl-tRNA synthetase, overview, the mutations confer a dominant lethal phenotype, overproduction of the inactive enzyme represses the expression of the wild-type chromosomal copy of the gene to an extent incompatible with bacterial growth, phenotypes, overview
R583H
-
site-directed mutagenesis, the mutant shows altered substrate specificity, overview
additional information
Q58597
construction of mutants consisting of catalytic or editing enzyme domains, overview
Y120A
-
site-directed mutagenesis, the mutation has no effect on the deacylation activity
additional information
Q980D1, -
construction of mutants consisting of catalytic or editing enzyme domains, overview, construction of mutant strain PBL205 with a disruption of gene thrS, i.e. SSO3004-3050
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant mutant M129K D-aminoacyl-tRNA deacylase-like domain from inclusion bodies after expression in Escherichia coli strain BL21(DE3) in 0.1 M phosphate buffer at pH 8.0 containing 6 M guanidinium hydrochloride and 10 mM Tris-HCl, followed by nickel affinity chromatography, the denatured protein is refolded stepwise by dialysis in 0.1 M phosphate buffer, pH 8.0, containing 10 mM Tris-HCl
-