Information on EC 6.1.1.7 - Alanine-tRNA ligase

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

EC NUMBER
COMMENTARY
6.1.1.7
-
RECOMMENDED NAME
GeneOntology No.
Alanine-tRNA ligase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + L-alanine + tRNAAla = AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla = AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
kinetic mechanism, AlaRS belongs to the class II of aminoacyl-tRNA ligases due to the position of aminoacylation on the 3'-terminal tRNA ribose, and the topology and tRNAbinding orientation of the active-site protein fold, class II synthetases are rate-limited by a step prior to aminoacyl transfer, the distinct mechanistic signatures of class I versus class II tRNA synthetases ensure rapid turnover of aminoacyl-tRNAs during protein synthesis
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Aminoacylation
-
-
-
-
Aminoacylation
-
-
Aminoacylation
-
-
Aminoacylation
-
-
Aminoacylation
Q54Y20
-
Aminoacylation
Q9U6B6
-
Aminoacylation
Q9U022
-
esterification
-
-
-
-
esterification
-
-
esterification
-
-
esterification
-
-
esterification
Q54Y20
-
esterification
Q9U6B6
-
esterification
Q9U022
-
PATHWAY
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
tRNA charging
-
SYSTEMATIC NAME
IUBMB Comments
L-Alanine:tRNAAla ligase (AMP-forming)
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ala-tRNA synthetase
-
-
-
-
Ala-tRNA synthetase
Q54Y20
-
Ala-tRNA synthetase
Q9U6B6
-
Ala-tRNA synthetase
-
-
Ala-tRNA synthetase
Q9U022
-
Ala-tRNA synthetase
-
-
Ala-tRNA synthetase
-
-
Alanine transfer RNA synthetase
-
-
-
-
Alanine transfer RNA synthetase
Q54Y20
-
Alanine transfer RNA synthetase
Q9U6B6
-
Alanine transfer RNA synthetase
-
-
Alanine transfer RNA synthetase
Q9U022
-
Alanine transfer RNA synthetase
-
-
Alanine transfer RNA synthetase
-
-
Alanine translase
-
-
-
-
Alanine translase
Q54Y20
-
Alanine translase
Q9U6B6
-
Alanine translase
-
-
Alanine translase
Q9U022
-
Alanine translase
-
-
Alanine translase
-
-
Alanine tRNA synthetase
-
-
-
-
Alanine tRNA synthetase
Q54Y20
-
Alanine tRNA synthetase
Q9U6B6
-
Alanine tRNA synthetase
-
-
Alanine tRNA synthetase
Q9U022
-
Alanine tRNA synthetase
-
-
Alanine tRNA synthetase
-
-
Alanine--tRNA ligase
-
-
-
-
Alanine-transfer RNA ligase
-
-
-
-
Alanine-transfer RNA ligase
Q54Y20
-
Alanine-transfer RNA ligase
Q9U6B6
-
Alanine-transfer RNA ligase
-
-
Alanine-transfer RNA ligase
Q9U022
-
Alanine-transfer RNA ligase
-
-
Alanine-transfer RNA ligase
-
-
alanine-tRNA ligase
Q54Y20
-
alanine-tRNA ligase
Q9U6B6
-
alanine-tRNA ligase
-
-
alanine-tRNA ligase
Q9U022
-
alanine-tRNA ligase
-
-
alanine-tRNA ligase
-
-
alanyl tRNA ligase
-
-
alanyl tRNA ligase
-
-
Alanyl-transfer ribonucleate synthetase
-
-
-
-
Alanyl-transfer ribonucleate synthetase
Q54Y20
-
Alanyl-transfer ribonucleate synthetase
Q9U6B6
-
Alanyl-transfer ribonucleate synthetase
-
-
Alanyl-transfer ribonucleate synthetase
Q9U022
-
Alanyl-transfer ribonucleate synthetase
-
-
Alanyl-transfer ribonucleate synthetase
-
-
Alanyl-transfer ribonucleic acid synthetase
-
-
-
-
Alanyl-transfer ribonucleic acid synthetase
Q54Y20
-
Alanyl-transfer ribonucleic acid synthetase
Q9U6B6
-
Alanyl-transfer ribonucleic acid synthetase
-
-
Alanyl-transfer ribonucleic acid synthetase
Q9U022
-
Alanyl-transfer ribonucleic acid synthetase
-
-
Alanyl-transfer ribonucleic acid synthetase
-
-
Alanyl-transfer RNA synthetase
-
-
-
-
Alanyl-transfer RNA synthetase
Q54Y20
-
Alanyl-transfer RNA synthetase
Q9U6B6
-
Alanyl-transfer RNA synthetase
-
-
Alanyl-transfer RNA synthetase
Q9U022
-
Alanyl-transfer RNA synthetase
-
-
Alanyl-transfer RNA synthetase
-
-
alanyl-tRNA ligase
-
-
alanyl-tRNA synthase
-
-
Alanyl-tRNA synthetase
-
-
-
-
Alanyl-tRNA synthetase
-
-
Alanyl-tRNA synthetase
Q54Y20
-
Alanyl-tRNA synthetase
Q9U6B6
-
Alanyl-tRNA synthetase
-
-
Alanyl-tRNA synthetase
Q9U022
-
Alanyl-tRNA synthetase
O58307
-
Alanyl-tRNA synthetase
Pyrococcus horikoshii OT-3
O58307
-
-
Alanyl-tRNA synthetase
-
-
Alanyl-tRNA synthetase
-
-
AlaRS
-
-
-
-
AlaRS
Q9U022
-
AlaRS
O58307
-
AlaRS
Pyrococcus horikoshii OT-3
O58307
-
-
Synthase, alanyl-transfer ribonucleate
-
-
-
-
Synthase, alanyl-transfer ribonucleate
Q54Y20
-
Synthase, alanyl-transfer ribonucleate
Q9U6B6
-
Synthase, alanyl-transfer ribonucleate
-
-
Synthase, alanyl-transfer ribonucleate
Q9U022
-
Synthase, alanyl-transfer ribonucleate
-
-
Synthase, alanyl-transfer ribonucleate
-
-
additional information
-
the enzyme belongs to the MurMN/Fem-ABX family of tRNA-dependent ligases
CAS REGISTRY NUMBER
COMMENTARY
9031-71-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
gene alaS
SwissProt
Manually annotated by BRENDA team
gene alaS is nuclear-encoded, mitochondrial isozyme
Q9U6B6
SwissProt
Manually annotated by BRENDA team
K12 strain KL386 that carries the gene on a recombinant pBR322 plasmid
-
-
Manually annotated by BRENDA team
overproducing strain
-
-
Manually annotated by BRENDA team
purified recombinant His-tagged enzyme
-
-
Manually annotated by BRENDA team
recombinant wild-type from overexpression in Escherichia coli
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes with replacement of Lys73 with Gln, Asn, Ala or Glu
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes with replacement ofcysteine residues, Cys76Ser, Cys290Ser, Cys412Ser, Cys665Ser
-
-
Manually annotated by BRENDA team
Escherichia coli overproducing
overproducing strain
-
-
Manually annotated by BRENDA team
expression in Pichia sp.
-
-
Manually annotated by BRENDA team
strain OT3, gene PH0574
SwissProt
Manually annotated by BRENDA team
Pyrococcus horikoshii OT-3
strain OT3, gene PH0574
SwissProt
Manually annotated by BRENDA team
mitochondrial and cytoplasmic isozymes encoded by a single nuclear gene ALA1
-
-
Manually annotated by BRENDA team
strain alphaS288C
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae alphaS288C
strain alphaS288C
-
-
Manually annotated by BRENDA team
strains 159, Pn16
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain HB8
-
-
Manually annotated by BRENDA team
strain HB8, overproduced in Escherichia coli
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
the relatively modest specificity of the AlaRS editing domain may provide a rationale for the widespread phylogenetic distribution of AlaX free-standing editing domains, thereby contributing a further mechanism to lower concentrations of misacylated tRNAAla
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + alanine + tRNAAla
?
show the reaction diagram
-
key role in protein biosynthesis
-
-
-
ATP + alanine + tRNAAla
?
show the reaction diagram
-
key role in protein biosynthesis
-
-
-
ATP + glycine + tRNAAla
AMP + diphosphate + glycyl-tRNAAla
show the reaction diagram
P00957
-
-
-
?
ATP + L-alanine + dual specific tRNAPhe
AMP + diphosphate + L-alanyl-tRNAPhe
show the reaction diagram
-
effects of deoxynucleotide substitutions in the tRNAPhe substrate on the enzyme activity, overview
-
?
ATP + L-alanine + dual specific tRNAPhe(17)
AMP + diphosphate + L-alanyl-tRNAPhe(17)
show the reaction diagram
-
site-specific modification leads to deoxynucleotide substituted yeast tRNA substrate, the backbone is interrupted on the 3'-side of nucleotide 17
-
?
ATP + L-alanine + dual specific tRNAPhe(38)
AMP + diphosphate + L-alanyl-tRNAPhe(38)
show the reaction diagram
-
site-specific modification leads to deoxynucleotide substituted yeast tRNA substrate, the backbone is interrupted on the 3'-side of nucleotide 38
-
?
ATP + L-alanine + dual specific tRNAPhe(57)
AMP + diphosphate + L-alanyl-tRNAPhe(57)
show the reaction diagram
-
site-specific modification leads to deoxynucleotide substituted yeast tRNA substrate, the backbone is interrupted on the 3'-side of nucleotide 57
-
?
ATP + L-alanine + tmRNA
AMP + diphosphate + L-alanyl-tmRNA
show the reaction diagram
-
recombinant transfer messenger RNA substrate from Thermus thermophilus expressed in Escherichia coli
-
?
ATP + L-alanine + tmRNA
AMP + diphosphate + L-alanyl-tmRNA
show the reaction diagram
-
transfer messenger RNA, 75fold reduced activity compared to cognate tRNAAla
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
P74941, -
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
P00957
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Q9U022
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Q54Y20
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Q9U6B6
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
O58307, -
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
marked specificity for ATP. ADP, GTP, ITP, CTP and UTP are inactive
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
2-step reaction
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
wild-type and mutant Escherichia coli tRNAAla substrates, enzyme interacts with 2'-hydroxyls in the acceptor stem of the tRNA substrate
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
a two-step reaction, class II synthetases are rate-limited by a step prior to aminoacyl transfer
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
O58307, -
AlaRS hydrolyzes mischarged tRNA by the catalytic zinc ion in the editing domain, modeling, overview
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
the aminoacylation reaction takes place in two steps catalyzed by the same active site: the synthesis of an aminoacyladenylate as an activated intermediate from the amino acid and ATP and the transfer of the aminoacyl moiety to the 3'-terminus of the cognate tRNA to yield the aminoacyl-tRNA, the synthetic active site of AlaRS misrecognizes noncognate glycine and serine as well as recognizing the cognate alanine and produces GlytRNAAla and Ser-tRNAAla, the editing domain hydrolyzes the incorrect products GlytRNAAla and Ser-tRNAAla and thus contributes to accurate aminoacylation, three tRNA isoacceptors, tRNAAla1, tRNA and tRNAAla3
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Escherichia coli overproducing
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Saccharomyces cerevisiae alphaS288C
-
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Pyrococcus horikoshii OT-3
O58307
-, AlaRS hydrolyzes mischarged tRNA by the catalytic zinc ion in the editing domain, modeling, overview
-
-
?
ATP + L-alanine + tRNAPyl
AMP + diphosphate + L-alanyl-tRNAPyl
show the reaction diagram
-
-
-
-
?
ATP + L-serine + tRNAAla
AMP + diphosphate + L-seryl-tRNAAla
show the reaction diagram
P00957
-
-
-
?
ATP + lipid II L-alanine + tRNAAla
?
show the reaction diagram
-
-
-
-
?
ATP + lipid II L-serine + tRNAAla
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
aminoacylates 9-base pair RNA duplexes whose sequences are based on the acceptor stems of either E. coli or human alanine tRNAs
-
-
-
additional information
?
-
-
synthesis of alanyl hydroxamate with high concentrations of hydroxylamine
-
-
-
additional information
?
-
P74941, -
alanine dependent ATP-diphosphate exchange: alanine + ATP + enzyme /alanine-AMP-enzyme + diphosphate
-
-
-
additional information
?
-
-
alanine dependent ATP-diphosphate exchange: alanine + ATP + enzyme /alanine-AMP-enzyme + diphosphate
-
-
-
additional information
?
-
-
alanine dependent ATP-diphosphate exchange: alanine + ATP + enzyme /alanine-AMP-enzyme + diphosphate
-
-
-
additional information
?
-
-
enzyme contains a second active site which prevents mistranslation (mistaking Ser or Gly for Ala) and which is specially designed for hydrolytic editing
-
-
-
additional information
?
-
-
analysis of the deacylation activities of the wild type and five different Escherichia coli AlaRS editing site substitution mutants using the free-standing Pyrococcus horikoshii AlaX editing domain complexed with serine as a model and both Ser-tRNAAla and Ala-tRNAAla as substrates, overview
-
-
-
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 + alanine + tRNAAla
?
show the reaction diagram
-
key role in protein biosynthesis
-
-
-
ATP + alanine + tRNAAla
?
show the reaction diagram
-
key role in protein biosynthesis
-
-
-
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
-
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Q9U022
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Q54Y20
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Q9U6B6
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
O58307, -
-
-
-
?
ATP + L-alanine + tRNAAla
AMP + diphosphate + L-alanyl-tRNAAla
show the reaction diagram
Pyrococcus horikoshii OT-3
O58307
-
-
-
?
additional information
?
-
-
analysis of the deacylation activities of the wild type and five different Escherichia coli AlaRS editing site substitution mutants using the free-standing Pyrococcus horikoshii AlaX editing domain complexed with serine as a model and both Ser-tRNAAla and Ala-tRNAAla as substrates, overview
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
-
or Mg2+ required
KCl
-
optimum concentration: 50-100 mM
Mg2+
-
optimum: 5-10 mM
Mg2+
-
or Co2+ required
Mg2+
-
6-16 mM
Zinc
-
thightly bound
Zn2+
-
removal of Zn2+ leads to over 50% reduced activity, required for conformational stability and flexibility important for the enzymes activity
Zn2+
O58307
active site bound by His9, His13, His120, and Cys116, involved in the catalytic reaction
Zn2+
-
potential role of the coordinated zinc in editing substrate specificity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
-
2-mercaptoethylamine
-
-
5,5'-dithiobis(2-nitrobenzoate)
-
-
5,5'-dithiobis(2-nitrobenzoate)
-
reversed by DTT
elongation factor Tu
-
strongly inhibits the aminoacylation of tmRNA, deviation from the mechanism of aminoacyl ester bond protection upon formation of the canonical ternary complx between the elongation gactor Tu, nucleotide cofactor and aminoacyl-tRNA
-
L-cysteine
-
-
L-glutathione
-
-
monobromobimane
-
-
p-Hydroxymercuriphenylsulfonic acid
-
-
p-Substituted mercuribenzoate
-
-
sulfhydryl group containing substances
-
-
Urea
-
denaturation at 3.3 M, the Zn2+-depleted enzyme is much more sensitive to denaturation by urea than the native enzyme
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2,6-dichlorophenol-indophenol
-
enhances activity
spermine
-
stimulates only at low Mg2+ concentrations
Activator macromolecule
-
260000-350000 MW, decreases Km values for ATP and Ala
-
additional information
-
elongation factor Tu has no stimulating effect on the enzyme with both substrates: tRNAAla and Ala-tmRNA, but it protects both acylated RNAs against hydrolysis of the alanyl ester bond
-
additional information
-
elongation factor Tu protects acylated tRNAAla against hydrolysis of the alanyl ester bond
-
additional information
-
the class II enzyme does not show tight product binding and formation of ternary complexes with EF-Tu, it is not activated by EF-Tu
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.23
-
alanine
-
Cys665Ser mutant
0.3
-
alanine
-
wild-type AlaRS and mutant AlaRS(K73Q), ATP-diphosphate exchange
0.31
-
alanine
-
mutant AlaRS(K73A), ATP-diphosphate exchange
0.33
-
alanine
-
mutant Cys76Ser
0.34
-
alanine
-
-
0.34
-
alanine
-
wild-type
0.47
-
alanine
-
Cys412Ser mutant
0.5
-
alanine
-
Cys290Ser mutant
1.41
-
alanine
-
mutant AlaRS(K73E), ATP-diphosphate exchange
0.083
-
ATP
-
wild-type enzyme
0.095
-
ATP
-
Cys665Ser mtant
0.12
-
ATP
-
Cys412Ser mutant
0.14
-
ATP
-
wild-type AlaRS and mutant AlaRS(K73E), AlaRS(K73Q), ATP-diphosphate exchange
0.18
-
ATP
-
Cys76Ser mutant
0.2
-
ATP
-
mutant AlaRS(K73A), ATP-diphosphate exchange
0.21
-
ATP
-
Cys290Ser mutant
0.3
-
ATP
-
-
1.7
-
ATP
-
-
26
-
glycine
P00957
wild-type, 22C, pH 8.0
42
-
glycine
P00957
mutant D235N, 22C, pH 8.0
88
-
glycine
P00957
mutant G237A, 22C, pH 8.0
493
-
glycine
P00957
mutant D235A, 22C, pH 8.0
0.05
-
L-alanine
P00957
wild-type, 22C, pH 8.0
0.8
-
L-alanine
-
-
73
-
L-alanine
P00957
mutant G237A, 22C, pH 8.0
178
-
L-alanine
P00957
mutant D235N, 22C, pH 8.0
502
-
L-alanine
P00957
mutant D235A, 22C, pH 8.0
31
-
L-serine
P00957
wild-type, 22C, pH 8.0
58
-
L-serine
P00957
mutant D235N, 22C, pH 8.0
70
-
L-serine
P00957
mutant G237A, 22C, pH 8.0
600
-
L-serine
P00957
above 600, mutant D235A, 22C, pH 8.0
0.0331
-
lipid II L-Ala
-
strain 159
0.04
-
lipid II L-Ala
-
strain Pn16
0.125
-
lipid II L-Ser
-
strain Pn16
0.146
-
lipid II L-Ser
-
strain 159
0.024
-
tmRNA
-
pH 7.6, 37C, recombinant His-tagged enzyme
-
0.023
-
tRNA
-
mutant AlaRS(K73E), aminoacylation
0.000104
-
tRNAAla
-
-
0.00028
-
tRNAAla
-
-
0.0011
-
tRNAAla
-
wild-type and mutant substrate from Escherichia coli, pH 7.5, 37C
0.0014
-
tRNAAla
-
-
0.0015
-
tRNAAla
-
pH 7.6, 37C, recombinant His-tagged enzyme
0.0024
-
tRNAAla
-
wild-type AlaRS, aminoacylation
0.022
-
tRNAAla
-
mutant AlaRS(K73Q), aminoacylation
0.028
-
tRNAAla
-
mutant AlaR(K73A), aminoacylation
0.0021
-
tRNAPhe
-
native dual specific tRNAPhe derivative substrate from Saccharomyces cerevisiae, pH 7.5, 37C
0.0019
-
tRNAPhe(17)
-
tRNAPhe mutant tRNAPhe(17), pH 7.5, 37C
-
0.0031
-
tRNAPhe(38)
-
tRNAPhe mutant tRNAPhe(38), pH 7.5, 37C
-
0.017
-
tRNAPhe(57)
-
tRNAPhe mutant tRNAPhe(57), pH 7.5, 37C
-
0.00063
-
liver tRNA
-
Bombyx mori tRNA
-
additional information
-
additional information
-
kinetics, ternary complex formation
-
additional information
-
additional information
-
kinetics
-
additional information
-
additional information
-
thermodynamics, pre-steady-state and single turnover kinetics, steady-state and transient kinetic analyses, recombinant His-tagged enzyme
-
additional information
-
additional information
-
steady-state kinetic parameters for deacylation of aa-tRNAAla at 37C and pH 7.5
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
79
-
ATP
-
Cys412Ser mutant
82
-
ATP
-
mutant AlaRS(K73Q), ATP-diphosphate exchange
84
-
ATP
-
wild-type
89
-
ATP
-
AlaRS, ATP-diphosphate exchange
89
-
ATP
-
Cys665Ser mutant
110
-
ATP
-
Cys290Ser mutant
120
-
ATP
-
AlaRS(K73A), ATP-diphosphate exchange
120
-
ATP
-
Cys76Ser mutant
0.0035
-
glycine
P00957
mutant D235A, 22C, pH 8.0
0.0058
-
glycine
P00957
mutant D235N, 22C, pH 8.0
1.6
-
glycine
P00957
mutant G237A, 22C, pH 8.0
3.7
-
glycine
P00957
wild-type, 22C, pH 8.0
0.0074
-
L-alanine
P00957
mutant D235N, 22C, pH 8.0
0.069
-
L-alanine
P00957
mutant D235A, 22C, pH 8.0
1.2
-
L-alanine
P00957
wild-type, 22C, pH 8.0
1.4
-
L-alanine
P00957
mutant G237A, 22C, pH 8.0
0.0028
-
L-serine
P00957
mutant G237A, 22C, pH 8.0
0.0046
-
L-serine
P00957
mutant D235N, 22C, pH 8.0
2.4
-
L-serine
P00957
wild-type, 22C, pH 8.0
0.0635
-
lipid II L-Ala
-
strain Pn16
0.071
-
lipid II L-Ala
-
strain 159
0.0166
-
lipid II L-Ser
-
strain 159; strain Pn16
0.043
-
tmRNA
-
pH 7.6, 37C, 0.001 mg recombinant His-tagged enzyme
-
0.1
-
tmRNA
-
pH 7.6, 50C
-
0.19
-
tRNAAla
-
mutant substrate from Escherichia coli, pH 7.5, 37C
0.2
-
tRNAAla
-
pH 7.6, 37C, 0.00025 mg recombinant His-tagged enzyme
0.24
-
tRNAAla
-
mutant AlaRS(K73E), aminoacylation
0.44
-
tRNAAla
-
wild-type substrate from Escherichia coli, pH 7.5, 37C
1.4
-
tRNAAla
-
mutant AlaRS(K73A), aminoacylation
1.5
-
tRNAAla
-
AlaRS, aminoacylation
1.7
-
tRNAAla
-
mutant AlaRS(K73Q), ATP-diphosphate exchange
2
-
tRNAAla
-
-
2.2
-
tRNAAla
-
pH 7.6, 50C
0.25
-
tRNAPhe
-
native dual specific tRNAPhe substrate from Saccharomyces cerevisiae, pH 7.5, 37C
0.25
-
tRNAPhe(17)
-
tRNAPhe mutant tRNAPhe(17), pH 7.5, 37C
-
0.46
-
tRNAPhe(38)
-
tRNAPhe mutant tRNAPhe(38), pH 7.5, 37C
-
0.12
-
tRNAPhe(57)
-
tRNAPhe mutant tRNAPhe(57), pH 7.5, 37C
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
additional information
-
the wild-type AlaRS editing domain deacylates Ser-tRNAAla with a kcat/Km of 66 mM/s, equivalent to a rate enhancement of 6000 over the rate of enzyme-independent deacylation but only 12.2fold greater than the rate with Ala-tRNAAla. While the E664A and T567G substitutions only minimally decrease kcat/Km, Q584H, I667E, and C666A AlaRS are more compromised in activity, with decreases in kcat/Km in the range of 6fold, 6.6fold, and 15fold. C666A AlaRS is 1.7fold more active on Ala-tRNAAla relative to Ser-tRNAAl. Deacylation rates of Ser-tRNAAla and Ala-tRNAAla in the absence of enzyme are determined by fitting the progress curves to equations describing a first-order decay
0
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0007
-
-
substrate: lipid II L-Ser, strain 159
0.0009
-
-
substrate: lipid II L-Ser, strain Pn16
0.0087
-
-
substrate: lipid II L-Ala, strain Pn16
0.011
-
-
substrate: lipid II L-Ala, strain 159
0.027
-
-
alanyl-tRNA synthesis
0.105
-
-
purified enzyme
0.114
-
-
diphosphate-ATP exchange
0.168
-
-
alanyl-tRNA synthetase activity
2.4
-
-
-
2.65
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.8
-
-
assay at
7.4
8.4
-
-
7.5
-
-
alanyl-tRNA synthesis
7.5
-
-
alanyl-tRNA synthesis
7.5
-
-
assay at
7.5
-
-
assay at
7.5
-
-
assay at
7.5
-
-
assay at, deacylation assay with aminoacylated tRNAAla as substrate
7.6
-
-
assay at
7.6
-
-
assay at
8.5
-
-
ATP-diphosphate exchange
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.4
7.5
-
6.4: about 60% of maximal activity, 7.5: optimum
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at, deacylation assay with aminoacylated tRNAAla as substrate
60
-
P74941
aminoacylation
65
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
10
37
-
independent of temperature in this range
25
75
-
half-maximal activity at 25C and 75C with substrate tRNAAla, a sharp 345fold drop of activity occurs at 70C, temperature profile
35
65
-
half-maximal activity at 35C and 65C with substrate tmRNA, a sharp 345fold drop of activity occurs at 40C, temperature profile
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
additional information
-
isozyme pattern of expression and localization, overview
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
very poor mitochondrial targeting activity of the ALA1 leader peptide, mapping of the mitochondrial targeting signal, overview
Manually annotated by BRENDA team
additional information
-
isozyme pattern of expression and localization, overview
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
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)
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 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)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
101000
121000
-
sucrose density gradient centrifugation
116000
-
-
gel filtration
131000
-
-
gel filtration
141000
-
-
gel filtration, cytoplasmic enzyme
151000
-
-
gel filtration
153000
-
-
gel filtration
162000
-
-
gel filtration, mitochondrial enzyme
179000
-
-
Zn2+-depleted enzyme, sedimentation coefficient and analytical ultracentrifugation
186000
-
-
calculation from sedimentation and diffusion coefficients
186000
-
-
native enzyme, sedimentation coefficient and analytical ultracentrifugation
195000
-
-
gel filtration
205000
-
P74941
size exclusion chromatography
360000
-
-
gel filtration
380000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
monomer
-
1 * 110000, SDS-PAGE
monomer
-
-
monomer
-
1 * 128000, SDS-PAGE
monomer
-
1 * 118000, denaturing electrophoresis
monomer
-
1 * 115000, SDS-PAGE, gel filtration in 6 M guanidine-HCl
monomer
-
1 * 96000, recombinant His-tagged enzyme, SDS-PAGE
monomer
O58307
1 * 18152, crystal structure analysis, two-domain structure consisting of seven antiparallel beta-sheets and six helices
monomer
Pyrococcus horikoshii OT-3
-
1 * 18152, crystal structure analysis, two-domain structure consisting of seven antiparallel beta-sheets and six helices
-
tetramer
-
4 * 95000, SDS-PAGE
tetramer
-
4 * 95000 (alpha), SDS-PAGE
tetramer
Escherichia coli overproducing
-
4 * 95000, SDS-PAGE
-
monomer
Saccharomyces cerevisiae alphaS288C
-
1 * 128000, SDS-PAGE
-
additional information
-
decameric association complex in which dimeric AlaRS is the predominant species at 25C
additional information
-
AlaRS consists of four parts: an N-terminal aminoacylation active-site domain, a tRNA-recognition module, an editing domain and a C-terminal oligomerization domain
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
453 amino acid catalytic fragment Aa-AlaRS453
-
cocrystal structures with Mg2+-ATP, L-alanine, glycine and L-serine with each separately
-
AlaRS-DELTAC, comprising the aminoacylation, tRNA-recognition, and editing domains, and AlaRS-C, comprising the dimerization domain. The aminoacylation/tRNA-recognition domains contain an insertion incompatible with the class-specific tRNA-binding mode. The editing domain is fixed tightly via hydrophobic interactions to the aminoacylation/tRNA-recognition domains, on the side opposite from that in threonyl-tRNA synthetase. A groove formed between the aminoacylation/tRNA-recognition domains and the editing domain appears to be an alternative tRNA-binding site. The amino acid residues required for the G3:U70 base pair recognition are mapped in this groove. The dimerization domain consists of helical and globular subdomains. The helical subdomain mediates dimerization by forming a helixloophelix zipper. The globular subdomain, which is important for the aminoacylation and editing activities, has a positively-charged face suitable for tRNA binding
-, O28029
purified recombinant full-length enzyme, N-terminal domain, and C-terminal domain, hanging-drop vapour-diffusion method, mixing of 0.001 ml of protein and reservoir solution, and equilibration against 0.5 ml of reservoir solution at 20C, for crystallization of the AlaRS-FLtRNAAla complex, tRNAAla is heated at 80C for 5 min and is gradually cooled to room temperature for refolding, in presence of 1 mM AMP-PNP, different methods for the different protein samples are used, overview, X-ray diffraction structure determination and analysis at 3.2-3.5 A resolution
-
molecular replacement with the apo-AlaRS structure from Aquifex aeolicus,Protein Data Bank accession code 1RIQ, as the initial model
P00957
of small fragments
-
monomer structures of C-terminally truncated AlaRS, with both activation and editing domains in the apo form, in complex with an Ala-AMP analog, and in a high-resolution lysine-methylated form. Docking of the editing domain to the activation domain occurs opposite from the predicted tRNA-binding surface. The editing site is positioned more than 35 A from the activation site. Results suggest that tRNA translocation via a canonical CCA flipping is unlikely to occur in AlaRS. Zinc binds in the editing site, in which the specific coordination of zinc will be facilitated by a conserved GGQ motif
O58035
purified recombinant wild-type and selenomethionine-labeld AlaRS editing-domain homolog, PH0574, 37.6 mg/ml protein, 27.5%w/v PEG 4000, and 100 mM MES-Na, pH 6.3, using a full-automatic protein crystallization and observation system, X-ray diffraction structure determination and analysis at 1.45 A resolution
O58307
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
65
-
P74941
stable up to
85
-
P74941
denaturation at
additional information
-
-
the Zn2+-depleted enzyme shows reduced viscosity at 10C compared to 37C
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
sensitive to repeated freezing and thawing
-
Zn2+ stabilizes the enzyme
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80C, 3 months, retains approximately 90% of activity
-
-20C, 50% glycerol, stable for at least 1 year
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant full-length AlaRS and AlaRS domains from Escherichia coli strain BL21 by anion exchange chromatography followed by hydrophobic interaction and adsorption chromatography, respectively
-
partial, from a multienzyme complex containing at least 16 aminoacyl-tRNA synthetase activities
-
purified by Ni-NTA chromatography and SDS gel electrophoresis
-
recombinant PH0574, from Escherichia coli strain BL21 by two steps of anion exchange chromatography, hydroxyapatite chromatography, and gel filtration
O58307
using Ni-Sepharose columns and anion-exchange chromatography. The yield of purified protein is 4-5 mg of protein per litre of culture, judged to be above 95% purity by SDS-PAGE
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
expression in Escherichia coli strain BL21
-
expression of the full-length AlaRS, of the N-terminal fragment lacking the C-terminal oligomerization domain, residues 1-739, and of the C-terminal oligomerization domain of AlaRS, residues 737-906, in Escherichia coli strain BL21
-
gene alaS, DNA sequence determination and analysis, phylogenetic origin
Q54Y20
deletion mutants with reduced aminoacylation efficiency, expressed in Escherichia coli strain BL21
-
gene alaS, DNA sequence determination and analysis, phylogenetic origin
Q9U6B6
expression in Escherichia coli
P00957
gene alaS, DNA sequence determination and analysis, phylogenetic origin
Q9U022
expression in Pichia sp.
-
temperature-sensitve mutant tsET12
Q8CFX8
gene PH0574, expression in Escherichia coli strain BL21
O58307
gene ALA1, DNA and amino acid sequence determination and analysis, mitochondrial and cytoplasmic isozymes are encoded by a single nuclear gene ALA1, through alternative use of inframe successive ACG triplets and a downstream AUG triplet, overview, despite participation of the non-AUG-initiated leader peptide in mitochondrial localization, the leader peptide per se cannot target a cytoplasmic passenger protein into mitochondria under normal conditions, functional mapping
-
gene cdc64, DNA sequence determination and analysis, gene maps to a locus between met7 and prt1 on chromosome XV, gene derepresses gcn4 expression, the cdc64-1 mutant strain can be complemented by expression of the wild-type enzyme
-
recombinant MurN is expressed and purified as MBP fusion protein in Escherichia coli
-
expression in Escherichia coli
P74941
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C290S
-
mutant enzymes with replacement of Cys residues, Cys76Ser, Cys290Ser, Cys412Ser, Cys665Ser. Mutation of Cys665 to serine induces a 120-fold decrease in catalytic efficiency
C665S
-
mutant enzymes with replacement of Cys residues, Cys76Ser, Cys290Ser, Cys412Ser, Cys665Ser. Mutation of Cys665 to serine induces a 120-fold decrease in catalytic efficiency
C666A
-
site-directed mutagenesis, the mutant shows reduced deacylation rates of tRNAAla compared to the wild-type enzyme
C666A/Q584H
-
aminoacylation activity is unchanged from that of wild-type. In contrast to the wild-type protein mutant protein mischarges Ser onto tRNAAla. Consistent with this mischarging, deacylation of Ser-tRNA Ala by the mutant protein is undetecable. Mutant protein is sensitive to high concentrations of serine; Serine toxicity, experienced by a strain harboring an C666A/Q584H editing-defective alanyl-tRNA synthetase mutant, is rescued by an AlaXp-encoding transgene from Methanosarcina mazei. AlaXp is a free-standing editing domain homolog of AlaRS. Rescue is dependent on amino acid residues in AlaXp that are needed for its in vitro catalytic activity
C76S
-
mutant enzymes with replacement of Cys residues, Cys76Ser, Cys290Ser, Cys412Ser, Cys665Ser. Mutation of Cys665 to serine induces a 120-fold decrease in catalytic efficiency
D235A
P00957
no improvement in the discrimination between alanine and serine
D235E
P00957
no improvement in the discrimination between alanine and serine
D235N
P00957
no improvement in the discrimination between alanine and serine
D235Q
P00957
no improvement in the discrimination between alanine and serine
DELTA1-437
-
mutant protein containing a deleted aminoacylation domain: mutant protein is fully active for clearance of Ser-tRNAAla but it is inactive deacylate Ser-tRNAAla
DELTA1-437/731-875
-
mutant protein containing a deleted aminoacylation domain: mutant protein is inactive for clearance of Ser-tRNAAla. Using RNA-binding assays, it is shown that the inactivity of the mutant correlates with a lack of binding of tRNAAla. However, at much higher concentrations, mutant is able of specifically deacylating misacylated tRNAAla. Thus, the catalytic site for editing is not disrupted instead, the reduction in editing activity results from a loss of affinity for tRNA
DELTA1-437/R693K
-
a region important for tRNA-specificity is further localized to a predicted strand-loop-strand motif within the region 438-875. Arg 693 is highly conserved. Mutant R693K has relaxed specificity for tRNAThr, and deacylated Ser-tRNAThr. Thus, the AlaRS editing domain shares a second, independent way to recognize tRNAAla
E664A
-
site-directed mutagenesis, the mutant shows reduced deacylation rates of tRNAAla compared to the wild-type enzyme
G237A
P00957
mutation G237A introduces bulk into the alanine-binding pocket, with little other change in the pocket or the surrounding atoms. Shrinking of the alanine-binding pocket sharply raises the Km for alanine but does not greatly perturb the Km for serine
I667E
-
site-directed mutagenesis, the mutant shows reduced deacylation rates of tRNAAla compared to the wild-type enzyme
L73A
-
mutant enzymes with replacement of Lys73 with Gln, Asn, Ala or Glu show reduction in catalytic efficiency in aminoacylation assay. Glu substitution causes a 5-fold decrease in affinity for alanine
L73E
-
mutant enzymes with replacement of Lys73 with Gln, Asn, Ala or Glu show reduction in catalytic efficiency in aminoacylation assay. Glu substitution causes a 5-fold decrease in affinity for alanine
L73N
-
mutant enzymes with replacement of Lys73 with Gln, Asn, Ala or Glu show reduction in catalytic efficiency in aminoacylation assay. Glu substitution causes a 5-fold decrease in affinity for alanine
Q584N
-
site-directed mutagenesis, the mutant shows reduced deacylation rates of tRNAAla compared to the wild-type enzyme
additional information
-, O28029
expression of truncated forms AlaRS-DELTAC, comprising the aminoacylation, tRNA-recognition, and editing domains, and AlaRS-C, comprising the dimerization domain
L73Q
-
mutant enzymes with replacement of Lys73 with Gln, Asn, Ala or Glu show reduction in catalytic efficiency in aminoacylation assay. Glu substitution causes a 5-fold decrease in affinity for alanine
additional information
-
engineering of the enzyme by introduction of a mutation that compensates for a deletion
additional information
-
K12 strain KL386 that carries the gene on a recombinant pBR322 plasmid
additional information
-
the needed tRNA interaction energy is further localized to non-specific RNA-binding determinants located in the region between amino acids 808 and 875
additional information
-
mutagenesis of the AlaRS editing active site
T567G
-
site-directed mutagenesis, the mutant shows reduced deacylation rates of tRNAAla compared to the wild-type enzyme
additional information
Escherichia coli overproducing
-
engineering of the enzyme by introduction of a mutation that compensates for a deletion
-
additional information
-
temperature-sensitive mutation cdc64-1 might affect charging of tRNAAla and thereby initiation of cell division, the mutation causes G1 arrest in yeast cells corresponding to a type II Start phenotype, the mutant strain can be complemented by expression of the wild-type enzyme
additional information
-
overexpression of the cytoplasmic isozyme enables it to overcome the compartmental barrier and function in the mitochondria as well, but deletion of as few as eight amino acid residues from its amino-terminus eliminates such a potential, the sequence upstream of the first in-frame AUG initiator not only carries an unusual initiation site, but also contributes to the pattern of protein expression and localization
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
molecular biology
-
the unique widespread distribution of the free-standing editing domain homolog AlaXp is most probably due to singular difficulties, for translation, poised by alanine
molecular biology
-
a combined action of ligases MurM and MurN is required in order to rationalise the high level of dipeptide cross-links in penicillin-resistant Streptococcus pneumoniae, with ligase MurM showing the major difference between penicillin-resistant and penicillin-sensitive strains