Information on EC 6.1.1.3 - threonine-tRNA ligase

New: Word Map on EC 6.1.1.3
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
6.1.1.3
-
RECOMMENDED NAME
GeneOntology No.
threonine-tRNA ligase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Aminoacylation
esterification
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
-
threonine metabolism
-
-
tRNA charging
-
-
SYSTEMATIC NAME
IUBMB Comments
L-threonine:tRNAThr ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY hide
9023-46-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Aeropyrum pernix DSM 11879
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
several strains, overview, gene thrS
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Escherichia coli overproducing
overproducing strain
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
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
10-70% less active than with L-threonine
-
?
ATP + L-isoleucine + tRNAIle
AMP + diphosphate + L-isoleucyl-tRNAIle
show the reaction diagram
ATP + L-serine + tRNASer
?
show the reaction diagram
-
yeast mitochondrial threonyl-tRNA synthetase MST1 lacks an editing domain and utilizes pre-transfer editing to discriminate against serine. MST1 misactivates serine and edits seryl adenylate (Ser-AMP) in the absence of the cognate tRNA. MST1 hydrolyzes 80% of misactivated Ser-AMP at a rate 4fold higher than that for the cognate threonyl adenylate (Thr-AMP) while releasing 20% of Ser-AMP into the solution.
-
-
?
ATP + L-serine + tRNASer
AMP + diphosphate + L-seryl-tRNASer
show the reaction diagram
ATP + L-serine + tRNAThr
AMP + diphosphate + L-seryl-tRNAThr
show the reaction diagram
ATP + L-threonine + tRNA1Thr
AMP + diphosphate + L-threonyl-tRNA1Thr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNA2Thr
AMP + diphosphate + L-threonyl-tRNA2Thr
show the reaction diagram
-
-
-
-
?
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
Formycin 5'-triphosphate + L-threonine + tRNAThr
?
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
-
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 + tRNASer
?
show the reaction diagram
-
yeast mitochondrial threonyl-tRNA synthetase MST1 lacks an editing domain and utilizes pre-transfer editing to discriminate against serine. MST1 misactivates serine and edits seryl adenylate (Ser-AMP) in the absence of the cognate tRNA. MST1 hydrolyzes 80% of misactivated Ser-AMP at a rate 4fold higher than that for the cognate threonyl adenylate (Thr-AMP) while releasing 20% of Ser-AMP into the solution.
-
-
?
ATP + L-threonine + tRNA1Thr
AMP + diphosphate + L-threonyl-tRNA1Thr
show the reaction diagram
-
-
-
-
?
ATP + L-threonine + tRNA2Thr
AMP + diphosphate + L-threonyl-tRNA2Thr
show the reaction diagram
-
-
-
-
?
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
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
NH4+
-
partial activation
Phosphorus
-
phosphoprotein containing phosphoserine, activity can be modulated by reversible phosphorylation
Rb+
-
partial activation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S,3R)-2,3-diamino-N-(((E)-3-(6-aminopyrimidin-4-yl)-styryl)sulfonyl)butanamide
(2S,3R)-2-amino-3-hydroxy-N-((3-(1-oxoisoindolin-5-yl)-phenyl)sulfonyl)butanamide
(2S,3R)-2-amino-3-hydroxy-N-((3-(3-methyl-1H-indazol-5-yl)phenyl)sulfonyl)butanamide
(2S,3R)-2-amino-3-hydroxy-N-((4-phenoxyphenyl)sulfonyl)-butanamide
(2S,3R)-2-amino-3-hydroxy-N-methyl-N-((3-(1-oxoisoindolin-5-yl)phenyl)sulfonyl)butanamide
(2S,3R)-2-amino-N'-(3-(4-amino-2-chloroquinazolin-7-yl)-phenyl)-3-hydroxybutanehydrazide
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxy-4-methylpentanamide
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxypentanamide
(2S,3R)-2-amino-N-((3-(1-amino-3-chloroisoquinolin-6-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-((3-(1-aminoisoquinolin-6-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-((3-(2,4-diaminoquinazolin-7-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-((3-(3-chloro-1H-indazol-5-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-((3-(4-amino-2-chloroquinazolin-7-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-((3-(4-amino-2-methylquinazolin-7-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-((3-(4-aminoquinazolin-7-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-((7-(6-aminopyrimidin-4-yl)naphthalen-2-yl)sulfonyl)-3-hydroxybutanamide
(2S,3R)-2-amino-N-(3-(4-amino-2-chloroquinazolin-7-yl)-benzyl)-3-hydroxybutanamide
(2S,3R)-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)sulfonyl)-2,3-dihydroxybutanamide
(2S,3R)-N-((7-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-2-amino-3-hydroxybutanamide
(2S,3R)-N-((7-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)naphthalen-2-yl)sulfonyl)-2-amino-3-hydroxybutanamide
2'-deoxyadenosine 5'-triphosphate
-
-
2'-O-Methyladenosine 5'-triphosphate
-
-
3'-Deoxyadenosine 5'-triphosphate
-
-
3'-O-Methyladenosine 5'-triphosphate
-
-
5'-O-[N-(threonyl)-sulfamoyl] adenosine
Borrelidin
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
operator mRNA domain 2
-
-
-
Purineriboside 5'-triphosphate
-
-
tert-butyl((2S,3R)-1-(3-(1H-indazol-5-yl)-benzenesulfonamido)-3-(tert-butoxy)-1-oxobutan-2-yl)-carbamate
threonyl-AMP
tubercidin 5'-triphosphate
-
-
Zn2+
-
inhibits the editing reaction
additional information
-
reactive oxygen species cause editing defect and misacylation by WT ThrRS
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
spermine
additional information
-
3-hydroxynorvaline enhances the ATPase function of the synthetic site, at a rate not increased by nonaminoacylatable tRNA
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.4
2-Chloroadenosine 5'-triphosphate
0.267 - 0.387
ATP
0.5
Formycin 5'-triphosphate
1.95 - 7
hydroxynorvaline
25 - 142
L-serine
0.00003 - 0.897
L-threonine
0.00189
threonine
-
-
0.00013 - 0.00095
tRNA1Thr
0.00027 - 0.0014
tRNA2Thr
0.00003 - 0.179
tRNAThr
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
42 - 90
ATP
21 - 22
hydroxynorvaline
1.3 - 30
L-serine
0.64 - 90
L-threonine
0.53
RNAThr
Escherichia coli
-
tRNAThr of E. coli
-
1.73
threonyl-tRNA
Saccharomyces carlsbergensis
-
-
0.033 - 0.118
tRNA1Thr
0.01 - 0.103
tRNA2Thr
0.00157 - 0.7
tRNAThr
0.037 - 0.23
tRNAThr of Thermus thermophilus
Thermus thermophilus
-
-
-
additional information
additional information
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.016
L-serine
Saccharomyces cerevisiae
-
in 100 mM Na-HEPES (pH 7.2), 30 mM KCl, 10 mM MgCl2, 2 mM potassium fluoride, at 37C
95
11.18
L-threonine
Saccharomyces cerevisiae
-
in 100 mM Na-HEPES (pH 7.2), 30 mM KCl, 10 mM MgCl2, 2 mM potassium fluoride, at 37C
250
1.093 - 171.7
tRNAThr
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.002262 - 0.05
(2S,3R)-2,3-diamino-N-(((E)-3-(6-aminopyrimidin-4-yl)-styryl)sulfonyl)butanamide
0.0000039 - 0.000091
(2S,3R)-2-amino-3-hydroxy-N-((3-(1-oxoisoindolin-5-yl)-phenyl)sulfonyl)butanamide
0.000083 - 0.00582
(2S,3R)-2-amino-3-hydroxy-N-((3-(3-methyl-1H-indazol-5-yl)phenyl)sulfonyl)butanamide
0.0331 - 0.193
(2S,3R)-2-amino-3-hydroxy-N-((4-phenoxyphenyl)sulfonyl)-butanamide
0.05
(2S,3R)-2-amino-3-hydroxy-N-methyl-N-((3-(1-oxoisoindolin-5-yl)phenyl)sulfonyl)butanamide
0.000034 - 0.000399
(2S,3R)-2-amino-N'-(3-(4-amino-2-chloroquinazolin-7-yl)-phenyl)-3-hydroxybutanehydrazide
0.0000225 - 0.000588
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxy-4-methylpentanamide
0.0000011 - 0.0000041
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxybutanamide
0.0000027 - 0.000032
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxypentanamide
0.00002 - 0.000463
(2S,3R)-2-amino-N-((3-(1-amino-3-chloroisoquinolin-6-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
0.000089 - 0.002242
(2S,3R)-2-amino-N-((3-(1-aminoisoquinolin-6-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
0.0000029 - 0.0000107
(2S,3R)-2-amino-N-((3-(2,4-diaminoquinazolin-7-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
0.000039 - 0.001518
(2S,3R)-2-amino-N-((3-(3-chloro-1H-indazol-5-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
0.0000003 - 0.05
(2S,3R)-2-amino-N-((3-(4-amino-2-chloroquinazolin-7-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
0.0000093 - 0.000072
(2S,3R)-2-amino-N-((3-(4-amino-2-methylquinazolin-7-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
0.0000009 - 0.0000033
(2S,3R)-2-amino-N-((3-(4-aminoquinazolin-7-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
0.0000007 - 0.000003
(2S,3R)-2-amino-N-((7-(6-aminopyrimidin-4-yl)naphthalen-2-yl)sulfonyl)-3-hydroxybutanamide
0.00014 - 0.000935
(2S,3R)-2-amino-N-(3-(4-amino-2-chloroquinazolin-7-yl)-benzyl)-3-hydroxybutanamide
0.05
(2S,3R)-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)sulfonyl)-2,3-dihydroxybutanamide
0.000004 - 0.0000095
(2S,3R)-N-((7-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-2-amino-3-hydroxybutanamide
0.0000006 - 0.0000024
(2S,3R)-N-((7-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)naphthalen-2-yl)sulfonyl)-2-amino-3-hydroxybutanamide
0.0000028 - 0.0000134
5'-O-[N-(threonyl)-sulfamoyl] adenosine
0.000004 - 0.006
Borrelidin
0.000132 - 0.05
tert-butyl((2S,3R)-1-(3-(1H-indazol-5-yl)-benzenesulfonamido)-3-(tert-butoxy)-1-oxobutan-2-yl)-carbamate
additional information
additional information
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
presteady-state, assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
presteady-state, assay at
50
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
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
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
association of the enzyme with high MW cellular component
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)
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)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
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)
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)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Staphylococcus aureus (strain MW2)
Staphylococcus aureus (strain MW2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
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
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
COMMENTARY hide
LITERATURE
homodimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
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
hanging drop vapor diffusion method, using 12% (w/v) PEG 4000, 21% (v/v) 2-methyl-2,4-pentanediol, and 0.1 M sodium citrate, pH 5.9
-
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
-
hanging drop vapor diffusion method, using 12% (w/v) PEG 4000, 21% (v/v) 2-methyl-2,4-pentanediol, and 0.1 M sodium citrate, pH 5.9
-
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
-
hanging drop vapor diffusion method, using 12% (w/v) PEG 4000, 21% (v/v) 2-methyl-2,4-pentanediol, and 0.1 M sodium citrate, pH 5.9
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
-
complexed with an analog of Ser-AMP, sitting drop vapor diffusion method, using 0.1 M Na2HPO4/KH2PO4 (pH 6.2), 0.2 M NaCl, 10% (w/v) PEG 8000
-
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
crystallized by the hanging-drop vapour diffusion method
-
enzyme overproduced in Escherichia coli
-
hanging drop vapor diffusion method, using 12% (w/v) PEG 4000, 21% (v/v) 2-methyl-2,4-pentanediol, and 0.1 M sodium citrate, pH 5.9
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
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
LITERATURE
-80C, about 30% loss of activity
-
-80C, stable for more than 18 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
5SrRNA-L5 protein complex containing both ThrRS and HisRS
-
metal chelate affinity column chromatography
Ni2+ affinity column chromatography
-
overproduced in Escherichia coli
-
recombinant from Escherichia coli