Information on EC 3.1.1.29 - aminoacyl-tRNA hydrolase

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

EC NUMBER
COMMENTARY hide
3.1.1.29
-
RECOMMENDED NAME
GeneOntology No.
aminoacyl-tRNA hydrolase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
N-substituted aminoacyl-tRNA + H2O = N-substituted amino acid + tRNA
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of carboxylic ester
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-asparagine biosynthesis III (tRNA-dependent)
-
-
L-cysteine biosynthesis II (tRNA-dependent)
-
-
SYSTEMATIC NAME
IUBMB Comments
aminoacyl-tRNA aminoacylhydrolase
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CAS REGISTRY NUMBER
COMMENTARY hide
9054-98-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
essential gene required for both vegetative growth and sporulation
-
-
Manually annotated by BRENDA team
MRE 600
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Wistar
-
-
Manually annotated by BRENDA team
Wistar
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
since build-up of peptidyl-tRNAs is toxic, defects in enzyme function result in cell death
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'(3')-O-L-(N,N-diacetyl-lysinyl)adenosine + H2O
?
show the reaction diagram
minimalist substrate
-
-
?
acetyl-histidine-tRNA + H2O
acetyl-histidine + tRNA
show the reaction diagram
-
-
-
?
acetyl-histidyl-tRNAHis + H2O
acetyl-histidine + tRNAHis
show the reaction diagram
-
-
-
-
?
Ala-tRNA + H2O
Ala + tRNA
show the reaction diagram
-
-
-
-
?
bulk peptidyl-tRNA + H2O
?
show the reaction diagram
D-tyrosine-tRNA + H2O
D-tyrosine + tRNA
show the reaction diagram
dephosphorylated diacetyl-lysine-tRNA + H2O
dephosphorylated diacetyl-lysine + tRNA
show the reaction diagram
-
-
-
?
dephosporylated formyl-methionine-tRNA + H2O
dephosphorylated formyl-methionine + tRNA
show the reaction diagram
-
-
-
?
diacetyl-Lys-tRNALys + H2O
diacetyl-Lys + tRNA
show the reaction diagram
diacetyl-lysine-tRNA + H2O
diacetyl-lysine + tRNA
show the reaction diagram
diacetyl-lysyl-tRNALys
diacetyl-Lys + tRNALys
show the reaction diagram
-
-
-
-
?
diacetyl-lysyl-tRNALys + H2O
diacetyl-lysine + tRNALys
show the reaction diagram
formyl-Met-tRNAfMet + H2O
formyl-Met + tRNAfMet
show the reaction diagram
-
-
-
-
?
formyl-methionine-tRNA + H2O
formyl-methionine + tRNA
show the reaction diagram
-
-
-
?
formyl-methionyl-tRNAfMet + H2O
formyl-methionine + tRNAfMet
show the reaction diagram
-
Escherichia coli formyl-methionyltRNAfMet, phosphorylated and dephosphorylated substrate
-
-
?
Glu-tRNA + H2O
Glu + tRNA
show the reaction diagram
-
-
-
-
?
Gly-tRNA + H2O
Gly + tRNA
show the reaction diagram
-
-
-
-
?
Gly-tRNAAla + H2O
Gly + tRNAAla
show the reaction diagram
-
-
-
?
L-Lys-tRNALys + H2O
L-lysine + tRNALys
show the reaction diagram
-
-
-
?
Leu-tRNA + H2O
Leu + tRNA
show the reaction diagram
Lys-tRNA + H2O
Lys + tRNA
show the reaction diagram
Met-tRNA + H2O
Met + tRNA
show the reaction diagram
-
-
-
-
?
Met-tRNAMet + H2O
Met + tRNAMet
show the reaction diagram
-
-
-
-
?
N-acetyl-Ala-tRNA + H2O
N-acetyl-Ala + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-Ala-tRNA(Ala) + H2O
N-acetyl-Ala + tRNA(Ala)
show the reaction diagram
-
-
-
-
?
N-acetyl-Glu-tRNA + H2O
N-acetyl-Glu + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-His-tRNA + H2O
N-acetyl-His + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-Leu-Gly-tRNA + H2O
N-acetyl-Leu-Gly + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-Leu-tRNA + H2O
N-acetyl-Leu + tRNA
show the reaction diagram
N-acetyl-Lys-tRNA + H2O
N-acetyl-Lys + tRNA
show the reaction diagram
N-acetyl-Met-tRNA + H2O
N-acetyl-Met + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-Phe-Phe-tRNA + H2O
N-acetyl-Phe-Phe + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-Phe-tRNA + H2O
N-acetyl-L-Phe + tRNA
show the reaction diagram
-
-
-
?
N-acetyl-Phe-tRNA + H2O
N-acetyl-Phe + tRNA
show the reaction diagram
N-acetyl-Phe-Val-tRNA + H2O
N-acetyl-Phe-Val + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-Ser-tRNA + H2O
N-acetyl-Ser + tRNA
show the reaction diagram
N-acetyl-Trp-tRNA + H2O
N-acetyl-Trp + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-Tyr-tRNA + H2O
N-acetyl-Tyr + tRNA
show the reaction diagram
-
-
-
-
?
N-acetyl-Val-tRNA + H2O
N-acetyl-Val + tRNA
show the reaction diagram
N-benzoyl-Gly-Gly-Phe-tRNA + H2O
N-benzoyl-Gly-Gly-Phe + tRNA
show the reaction diagram
-
-
-
-
?
N-benzoyl-Gly-GlyGly-Phe-tRNA + H2O
N-benzoyl-Gly-Gly-Gly-Phe + tRNA
show the reaction diagram
-
-
-
-
?
N-carbobenzyloxy-Phe-tRNA + H2O
N-carbobenzyloxy-Phe + tRNA
show the reaction diagram
-
-
-
-
?
N-formyl-Val-tRNA + H2O
N-formyl-Val + tRNA
show the reaction diagram
N-substituted aminoacyl-tRNA + H2O
N-substituted amino acid + tRNA
show the reaction diagram
oligolysyl-tRNA + H2O
oligo-Lys + tRNA
show the reaction diagram
-
-
-
-
?
Oregon Green-methionine-tRNA + H2O
Oregon Green-methionine + tRNA
show the reaction diagram
-
-
-
?
peptidyl-tRNA + H2O
?
show the reaction diagram
peptidyl-tRNA + H2O
peptide + tRNA
show the reaction diagram
peptidyl-tRNA hydrolase cleaves the ester bond between tRNA and the attached peptide in peptidyl-tRNA in order to avoid the toxicity resulting from its accumulation and to free the tRNA available for further rounds in protein synthesis
-
-
?
peptidyl-tRNAL + H2O
peptide + tRNA
show the reaction diagram
peptidyl-tRNALys + H2O
peptide + tRNALys
show the reaction diagram
Phe-Phe-tRNA + H2O
Phe-Phe + tRNA
show the reaction diagram
-
-
-
-
?
Phe-tRNA + H2O
Phe + tRNA
show the reaction diagram
phenyllactyl-Phe-tRNA + H2O
phenyllactyl-Phe + tRNA
show the reaction diagram
-
-
-
-
?
poly-Val-tRNA + H2O
poly-Val + tRNA
show the reaction diagram
-
-
-
-
?
Ser-tRNA + H2O
Ser + tRNA
show the reaction diagram
Ser-tRNAAla + H2O
Ser + tRNAAla
show the reaction diagram
-
-
-
?
Tyr-tRNA + H2O
Tyr + tRNA
show the reaction diagram
-
-
-
-
?
Val-tRNA + H2O
Val + tRNA
show the reaction diagram
Val-tRNAVal + H2O
Val + tRNAVal
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
diacetyl-lysyl-tRNALys + H2O
diacetyl-lysine + tRNALys
show the reaction diagram
N-substituted aminoacyl-tRNA + H2O
N-substituted amino acid + tRNA
show the reaction diagram
peptidyl-tRNA + H2O
peptide + tRNA
show the reaction diagram
P9WHN7
peptidyl-tRNA hydrolase cleaves the ester bond between tRNA and the attached peptide in peptidyl-tRNA in order to avoid the toxicity resulting from its accumulation and to free the tRNA available for further rounds in protein synthesis
-
-
?
peptidyl-tRNAL + H2O
peptide + tRNA
show the reaction diagram
-
Pth is a key protein at the crossroads to the function of several translational factors, accumulation of peptidyl-tRNA in the cells leads to depletion of aminoacyl-tRNA pools and halts protein biosynthesis, it is vital for cells to maintain Pth activity to deal with the pollution of peptidyl-tRNAs generated during the initiation, elongation and termination steps of protein biosynthesis, overview
-
-
?
peptidyl-tRNALys + H2O
peptide + tRNALys
show the reaction diagram
-
accumulation of peptidyl-tRNA due to enzyme misfunction is toxic to the cells, overproduction of tRNALys suppresses the effects of pthTs mutation at 41°C but not at 43°C, and increases the levels of aminoacyl-tRNA
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-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
activation is 25% lower than with Mg2+ or Mn2+
K+
-
0.3 mM, 77fold activation
KCl
-
300 mM, 77fold activation; activates about 80fold, optimally at 300 mM
MgCl2
-
40 mM, approx. 88fold activation
Zn2+
a zinc ion is coordinated by the conserved zinc-binding cluster in the C-domain, which is expected to be the enzymatic active site
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3'-deoxy-N[(O-methyl-L-tyrosyl)amino]adenosine
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3'-L-(N,N-diacetyl-lysinyl)amino-3'-deoxyadenosine
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-
70 S ribosome
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free N-carbobenzyloxy-Phe-tRNA is rapidly cleaved by the enzyme. When bound to a 30 S ribosome in the presence of poly(U), the substrate is hydrolyzed rapidly as when free. The addition of 50 S ribosomal subunits to form the 70S ribosomal binding complex protects the bound substrate from the enzyme
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deacetylated tRNA
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diethyldicarbonate
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0.5 mM, 90% inactivation after 10 min, activity can be recovered to 41% of initial activity by treatment wit 200 mM hydroxylamine
non-esterified tRNALys
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-
-
tRNA-formyl-methionine
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from Escherichia coli
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tRNA-lysine
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from Escherichia coli
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tRNAfMet
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; tRNAfMet from Escherichia coli
tRNALys
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tRNALys from Escherichia coli
Uncharged tRNA
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e.g. from Escherichia coli, at concentrations of 0.01 mM or above
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
spermidine
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0.1 mM spermidine HCl, 73fold activation
spermidine-HCl
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0.1 mM, 73fold activation; activates about 80fold, optimally at 0.1 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000016
acetyl-His-tRNAHis
-
pH 7.5, 50°C
0.000016
acetyl-histidyl-tRNA
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50°C, pH 7.5
0.000016
acetyl-histidyl-tRNAHis
-
pH 7.5, 50°C, recombinant enzyme
0.008
bulk peptidyl-tRNA
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at pH 5.0 and 25°C
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0.000003
dephosphorylated diacyl-lysine-tRNA
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50°C, pH 7.5
-
0.00003
dephosphorylated formyl-methioninyl-tRNA
-
50°C, pH 7.5
-
0.0000028 - 0.024
diacetyl-Lys-tRNALys
-
0.000011 - 0.006
diacetyl-lysine-tRNA
-
0.0000028 - 12.8
diacetyl-lysyl-tRNALys
0.000012
formyl-Met-tRNAfMet
-
pH 7.5, 50°C
0.00003
formyl-Met-tRNAfMet, dephosphorylated
-
pH 7.5, 50°C, dephosphorylated formyl-Met-tRNAfMet
-
0.000012
formyl-methionyl-tRNA
-
50°C, pH 7.5
0.000012 - 0.00003
formyl-methionyl-tRNAfMet
0.00471 - 0.0269
N-acetyl-Ala-tRNA(Ala)
-
0.0022
N-acetyl-Met-tRNA
-
pH 7.0, 37°C
0.0008
N-acetyl-Phe-tRNA
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pH 7.5, 30°C
0.000001
N-acetyl-phenylalanyl-tRNA
-
pH 7.0, 30°C
0.0055
Oregon Green-methionine-tRNA
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25°C, pH 7.5
-
0.0008
Phe-tRNA
-
pH 7.5, 30°C
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.4
acetyl-His-tRNA
Sulfolobus solfataricus
-
pH 7.5, 50°C
3.4
acetyl-histidyl-tRNA
Sulfolobus solfataricus
-
50°C, pH 7.5
3.4
acetyl-histidyl-tRNAHis
Sulfolobus solfataricus
-
pH 7.5, 50°C, recombinant enzyme
0.86 - 6.08
dephosphorylated diacyl-lysine-tRNA
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3
dephosphorylated formyl-methioninyl-tRNA
Sulfolobus solfataricus
-
50°C, pH 7.5
-
1.3
diacetyl-Lys-tRNA
Homo sapiens
Q9Y3E5
pH 7.5, 27°C
0.028 - 3.6
diacetyl-Lys-tRNALys
-
1.3 - 3.6
diacetyl-lysine-tRNA
-
0.002 - 2.2
diacetyl-lysyl-tRNALys
3
formyl-Met-tRNAfMet
Sulfolobus solfataricus
-
pH 7.5, 50°C
3
formyl-methionyl-tRNA
Sulfolobus solfataricus
-
50°C, pH 7.5
3
formyl-methionyl-tRNAfMet
Sulfolobus solfataricus
-
phosphorylated and dephosphorylated substrate, pH 7.5, 50°C, recombinant enzyme
5.8 - 11.7
N-acetyl-Ala-tRNA(Ala)
-
0.7
N-acetyl-Met-tRNA
Escherichia coli
-
pH 7.0, 37°C
3
o-hydroxycinnamic acid
Sulfolobus solfataricus
-
50°C, pH 7.5
9.3
Oregon Green-methionine-tRNA
Escherichia coli
-
25°C, pH 7.5
-
additional information
additional information
Escherichia coli
-
effect of mutations altering the 1-72 pair of E. coli tRNAMetf on the turnover-number
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.51
2'(3')-O-L-(N,N-diacetyl-lysinyl)adenosine
Escherichia coli
-
wild type enzyme, 28°C, 20 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 0.1 mM EDTA, 0.1 mM dithiothreitol
166174
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
12
3'-L-(N,N-diacetyl-lysinyl)amino-3'-deoxyadenosine
-
at pH 6.0, with 50 mM sodium acetate and 200 mM NaCl, at 28°C
0.00005
non-esterified tRNALys
-
pH 7.5, 50°C, recombinant enzyme
-
0.000095
tRNA
-
pH 7.5, 50°C, unfractionated Escherichia coli tRNA; unfractionated tRNA from Escherichia coli, 50°C, pH 7.5
0.0001
tRNA-formyl-methionine
-
tRNA-ormyl-methionine from Escherichia coli, 50°C, pH 7.5
-
0.00005
tRNA-lysine
-
tRNA-lysine from Escherichia coli, 50°C, pH 7.5
-
0.00011
tRNAfMet
-
pH 7.5, 50°C, recombinant enzyme
0.00011
tRNAformylMet
-
pH 7.5, 50°C, tRNAformylMet from Escherichia coli
-
0.00005
tRNALys
-
pH 7.5, 50°C, tRNALys from Escherichia coli
0.0001
Uncharged tRNA
-
pH 7.5, 50°C, recombinant enzyme
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0149
-
-
0.078
-
substrate D-tyrosyl-tRNA, 37°C, pH 7.5
0.15
-
substrae D-tyrosyl-tRNA
4.14
-
; purified recombinant enzyme
30
-
substrate diacetyl-lysine-tRNA
31.8
-
substrate diacetyl-lysine tRNA, 37°C, pH 7.5
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.5
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.5
approx. 38% at pH 6.0, approx. 20% of maximal activity at pH 8.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
enzyme from encysted embryos is specific for acetyl-Phe-tRNA. An unspecific hydrolase active on several N-substituted amiinoacyl-tRNAs is practically absent in the encysted embryos and during embryogenesis and appears abruptly during larval development
Manually annotated by BRENDA team
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paraproerythroblast, low activity
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
low activity
Manually annotated by BRENDA team
-
low activity
Manually annotated by BRENDA team
additional information
-
mutant strain AA7852 with temperature-sensitive Pth grown at 32°C and at 41°C
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Acinetobacter baumannii (strain ATCC 19606 / DSM 30007 / CIP 70.34 / JCM 6841 / NBRC 109757 / NCIMB 12457 / NCTC 12156 / 81)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Burkholderia thailandensis (strain ATCC 700388 / DSM 13276 / CIP 106301 / E264)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Francisella tularensis subsp. tularensis (strain SCHU S4 / Schu 4)
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Salmonella typhimurium (strain 14028s / SGSC 2262)
Streptococcus pyogenes serotype M49 (strain NZ131)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Vibrio cholerae serotype O1 (strain ATCC 39541 / Classical Ogawa 395 / O395)
Vibrio cholerae serotype O1 (strain M66-2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
13100
-
2 * 13100, deduced from nucleotide sequence
13500
-
x * 13500, SDS-PAGE
15000
-
2 * 15000, recombinant enzyme, SDS-PAGE; x * 15000, SDS-PAGE
20000
-
x * 20000, SDS-PAGE
20460
calculated from amino acid sequence
21000
-
1 * 21000, SDS-PAGE
21978
-
x * 21978, sequence calculation
24000
x * 24000, SDS-PAGE
46000
-
gel filtration
55000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme and in complex with cytidine or uridine, hanging drop vapor diffusion method, using 25% (w/v) PEG 10000, 0.3 M MgCl2 and 0.1 M HEPES buffer at pH 6.0
crystal structure analysis
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crystal structure at 1.3 A resolution
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crystallization by using polyethylene glycol as precipitant, recombinant enzyme
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crystallization using polyethylene glycol as precipitant and isopropanol as additive, crystal structure at 1.2 A
in complex with the tRNA CCA-acceptor-TpsiC domain of tRNAAla, sitting drop vapor diffusion method, using 100 mM sodium acetate buffer (pH 5.2), 20% (w/v) 1,4-butanediol and 30 mM glycyl-glycylglycine, at 20°C
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recombinant enzyme, sitting drop vapor diffusion techniques; sitting-drop vapor diffusion at room temperature, 10 mg/ml protein in 100 mM HEPES, pH 7.5 and 20% polyethylene glycol 10000 is equilibrated against a reservoir of the same solution, crystals diffract to 2.0 A
hanging drop vapor diffusion method, using 30% (w/v) PEG-1500 and 10% (v/v) isopropanol in 100 mM HEPES buffer, pH 6.5
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microbatch-under-oil method, using 0.1 M HEPES pH 7.5, 15% (w/v) PEG 8000, 5% (v/v) isopropanol or 0.1 M HEPES pH 7.5 and 5% (v/v) dioxane with 25% (w/v) PEG 8000
purified recombinant enzyme, X-ray diffraction structure determination and analysis at 1.98 A, 2.35 A, and 2.49 A resolution, molecular replacement
purified recombinant His-tagged enzyme, microbatch method, 0.002 ml each of the protein solution, containing 6 mg/ml protein, 20 mM Tris-HCl, pH 7.5, 100 mM NaCl and 2 mM 2-mercaptoethanol, and the precipitant solution, containing 20% w/v PEG 8000 in 0.1 M HEPES, pH 7.5, and 5% v/v 2-propanol or dioxane or 25% w/v PEG 8000, 100 mM sodium cacodylate, pH 6.6, and 5% v/v 2-propanol, are mixed, 5 days to 2 weeks, X-ray diffraction structure determination and analysis at 1.97-2.49 A resolution
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native enzyme and in complex with 3'-deoxy-N[(O-methyl-L-tyrosyl)amino]adenosine or 5-azacytidine, hanging drop vapor diffusion method, using 25% (w/v) PEG 4000, 5% (v/v) propan-2-ol and 0.1 M HEPES buffer, pH 7.5
hanging-drop vapour-diffusion method at 20°C, crystal structure is determined at 2.7 A resolution
hanging drop vapor diffusion method, using 0.03 M citric acid, 0.05 M Bis-Tris propane, 1% (v/v) glycerol, 3% (w/v) sucrose, 25% (w/v) PEG 6000 pH 7.6
D0ZJ57
hanging drop vapour diffusion method, 1.8 A resolution
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purified recombinant enzyme, hanging drop vapour diffusion method, 24°C, 0.0027 ml of 1.3 mg/ml protein in 20 mM Tris-HCl, pH 7.0, 0.1 mM EDTA, and 10 mM 2-mercaptoethanol are mixed with 0.0007 ml of 11 mg/ml tRNAfMet solution and 0.002 ml of reservoir solution containing 0.8 M LiSO4 and 1.6% PEG 8000, crystallization of tRNA free crystals within a few days, X-ray diffraction structure determination and analysis of native and HgBr2-containing crystals at 1.8-3.0 A resolution, modeling
sitting drop vapor diffusion method, using 100 mM phosphate-citrate buffer pH 4.2, and 50% (w/v) 2-methyl-2,4-pentanediol
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7.5
-
4°C, 24 h, stable
648113
6 - 8
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30°C, 30 min, stable
648114
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
10 min, 70% loss of activity
50
-
10 min, 85% loss of activity
additional information
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thermally-induced unfolding curve for MtPth indicates a simple two-state unfolding process without any intermediates, thermodynamic stability of the enzyme, pH 6.5, 25°C, overview
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
urea/guanidinium chloride-induced unfolding curve for MtPth indicates a simple two-state unfolding process without any intermediates, pH 6.5, 25°C
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10°C, 50% glycerol, purified enzyme is stable for several months
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-10°C, in presence of 1 mg/ml bovine serum albumin, no loss of activity
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-20°C, buffer solution: 0.05 M Tris-HCl, pH 7.0, 0.01 M 2-mercaptoethanol, 0.01 M magnesium acetate, or as an ammonium sulfate precipitate, 2 months without significant loss of activity
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22°C, 20 mM Bis-Tris pH 6.6, 50 mM NaCl, 2 mM dithiothreitol, several weeks, without loss of activity or precipitation
D0ZJ57
22°C, 20 mM sodium acetate, 150 mM sodium chloride, 2 mM dithiothreitol, pH 5.0, several months, minimal loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; recombinant PTH, heat, ammonium sulfate, Superdex 75, HI-Propyl, SP-Sepharose; recomninant PTH from Escherichia coli strain XL 1-Blue 8500fold by ultracentrifugation, heat treatment, ammonium sulfate fractionation, gel filtration, hydrophobic interaction chromatography, and ion exchange chromatography to homogeneity
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ammonium sulfate precipitation
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HisTrap column chromatography, Ni-NTA agarose column chromatography, and Q Sepharose column chromatography
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HisTrap nickel affinity column chromatography and Superdex 75 gel filtration
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Ni-chelating column chromatography
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Ni-NTA affinity resin column chromatography and Superdex G50 gel filtration
Ni-NTA column chromatography and Superdex G-75 gel filtration
Ni2+-chelating affinity column chromatography and Superose 12 gel filtration
D0ZJ57
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography
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recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis
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recombinant His-tagged Pth
recombinant Pth
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Super Q Toyopearl 650M column chromatography, Resource Q column chromatography, and Superdex 75 gel filtration
wild-type and mutant enzymes
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wild-type and mutant enzymes N10A, H20A, M67A, F66A, D93A, H113A, K142A
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as GST-fusion protein in Escherichia coli Rosetta pLysS (GST later cleaved), or in human HEK293T cells as FLAG-tagged protein
expressed in Escherichia coli BL21 (DE3) pMGK cells with eight nonnative residues at the C-terminus (LEHHHHHH) to facilitate protein purification
expressed in Escherichia coli BL21 pLysS cells
D0ZJ57
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3)pLysS cells
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expressed in Escherichia coli BL21(lambdaDE3) cells
expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL cells
expressed in Escherichia coli strain K37DELTArecAlambdaDE3
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expression in Escherichia coli
expression in Escherichia coli; full length human Pth2 clone as tempolate , subcloned into the NcoI/BamHI sites of the pET15b bacterial expression vector. The ligated plasmid is transfoirmed into Escherichia coli BL21(DE3) strain
expression in Escherichia coli; gene SS00175, DNA and amino acid sequence determination and analysis, complementation of an enzyme-deficient Escherichia coli mutant strain, and of two Saccharomyces cerevisiae gene YHR189w or YBL057c disruption mutants, overview, expression of PTH in Escherichia coli strain XL 1-Blue
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gene pth, orf Rv1014c, DNA and amino acid sequence determination and analysis, expression of wild-type and mutant C-terminally His-tagged enzymes in Escherichia coli strain BL21(DE3) and in Escherichia coli thermosensitive strain AA7852, the mutant strain is able to grow at the nonpermissive temperature of 42°C, at 39°C, overexpression of MtPth in AA7852 cells allowed the cells to remain viable in the presence of up to 200 mg/ml erythromycin, overview
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gene pth, orf Rv1014c, DNA and amino acid sequence determination, expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3)
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pth2 gene, expression in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D93A
turnover-number is 0.1% of the turnover-number for wild-type enzyme, Km-value for diacetyl-Lys-tRNALys is 1.67fold higher than the Km-value for the wild-type enzyme
D93N
-
4% of wild-type kcat
F66A
turnover-number is 26% of the turnover-number for wild-type enzyme, Km-value for diacetyl-Lys-tRNALys is 1.15fold higher than the Km-value for the wild-type enzyme
H113A
turnover-number is 33% of the turnover-number for wild-type enzyme, Km-value for diacetyl-Lys-tRNALys is 1.46fold higher than the Km-value for the wild-type enzyme
H188A
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the mutation results in a 5.4fold decrease in the kcat/Km value compared to the wild type enzyme
H20N
-
no activity
H20Q
-
no activity
K103Q
-
54% of wild-type kcat
K103R
-
68% of wild-type kcat
K103S
-
28% of wild-type kcat
K105Q
-
20% of wild-type kcat
K105R
-
26% of wild-type kcat
K105S
-
16% of wild-type kcat
K113Q
-
98% of wild-type kacat
K142A
turnover-number is 24% of the turnover-number for wild-type enzyme, Km-value for diacetyl-Lys-tRNALys is 4fold higher than the Km-value for the wild-type enzyme
M67A
turnover-number is 4.7% of the turnover-number for wild-type enzyme, Km-value for diacetyl-Lys-tRNALys is 70% of the Km-value for the wild-type enzyme
M67E
-
0.5% of wild-type kcat
N185A
-
the mutation results in a 5.7fold decrease in the kcat/Km value compared to the wild type enzyme
N185A/H188A
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the mutation results in a 7.7fold decrease in the kcat/Km value compared to the wild type enzyme
C166A
-
site-directed mutagenesis, the mutant effectively complements the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C
C67S
-
site-directed mutagenesis, the mutant effectively complements the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C
D95N
-
site-directed mutagenesis, the catalytic residue mutant is not able to complement the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C
H22N
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site-directed mutagenesis, the catalytic residue mutant is not able to complement the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C; site-directed mutagenesis, the mutation affects the enzyme structure, overview
N12D
-
site-directed mutagenesis, the catalytic residue mutant is not able to complement the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C
C166A
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site-directed mutagenesis, the mutant effectively complements the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C
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C67S
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site-directed mutagenesis, the mutant effectively complements the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C
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D95N
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site-directed mutagenesis, the catalytic residue mutant is not able to complement the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C
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H22N
-
site-directed mutagenesis, the catalytic residue mutant is not able to complement the enzyme-defective thermosensitive Escherichia coli mutant strain AA7852 for growth at 42°C; site-directed mutagenesis, the mutation affects the enzyme structure, overview
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C703A
mutation abolishes editing activity
D86A
-
kcat/Km for diacetyl-lysyl-tRNALys is 0.22% of wild-type value
D86A/K18A
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kcat/Km for diacetyl-lysyl-tRNALys is 0.17% of wild-type value
H25A
-
kcat/Km for diacetyl-lysyl-tRNALys is 13% of wild-type value
K118A
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kcat/Km for diacetyl-lysyl-tRNALys is 87% of wild-type value
K18A
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kcat/Km for diacetyl-lysyl-tRNALys is 0.2% of wild-type value
K56A
-
kcat/Km for diacetyl-lysyl-tRNALys is 12% of wild-type value
Q22A
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kcat/Km for diacetyl-lysyl-tRNALys is 20% of wild-type value
Q54A
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kcat/Km for diacetyl-lysyl-tRNALys is 47% of wild-type value
T90A
-
kcat/Km for diacetyl-lysyl-tRNALys is 1.4% of wild-type value
T98A
-
kcat/Km for diacetyl-lysyl-tRNALys is 34% of wild-type value
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
-
Pth is a potential drug target to control eubacterial infections
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