Information on EC 6.1.1.12 - Aspartate-tRNA ligase

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

EC NUMBER
COMMENTARY
6.1.1.12
-
RECOMMENDED NAME
GeneOntology No.
Aspartate-tRNA ligase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
conserved amino acid residues of functional importance in consensus motifs: Pro227 in motif 1, Arg332 and Glu334 in motif 2, Gly675, Gly679, Asp681, Arg682, and Pro700 in motif 3; conserved amino acid residues of functional importance in consensus motifs: Pro227 in motif 1, Arg332 and Glu334 in motif 2, Gly675, Gly679, Glu681, Arg682, and Pro700 in motif 3
P36419, Q9LCY8
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
active site residues
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
determination of structural features for discriminating or nondiscriminating aminoacylation activity
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
mechanism, active site structure, substrate binding, simulations of binding and free energy, protein-ligand modeling
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
mechanism, active site structure, substrate binding, simulations of binding and free energy
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
mechanism
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
pathway for tRNA binding and recognition is proposed
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
active site structure
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Asn binding sites K198 and H449, binding mechanism and free energies
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
RNA-binding motif XSKXXLKKXK
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
protein-RNA recognition mechanism, backbone interactions are an important functional component of the tRNA synthetase interaction
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
active site structure, ligand and substrate binding, and reaction mechanism
-
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
active site structure, AspRS substrate recognition and chiral specificity of wild-type enzyme and mutant Q199E, substrate binding in regular versus inverted orientation, overview
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
aminoacyl group transfer
-
-
aminoacyl group transfer
-
-
aminoacyl group transfer
-
-
Aminoacylation
-
-
-
-
Aminoacylation
-
-
Aminoacylation
-
-
Aminoacylation
-
-
Aminoacylation
-
-
Aminoacylation
Q38CB4, Q585M3, -
;
Aminoacylation
-, Q8I2B1
-
Aminoacylation
-
-
Aminoacylation
-, Q3U987
-
Aminoacylation
C4LZN0, -
-
esterification
-
-
-
-
esterification
-
-
esterification
-
-
esterification
-
-
esterification
-
-
esterification
Q38CB4, Q585M3, -
;
esterification
-, Q8I2B1
-
esterification
-
-
esterification
-, Q3U987
-
esterification
C4LZN0, -
-
PATHWAY
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
tRNA charging
-
SYSTEMATIC NAME
IUBMB Comments
L-Aspartate:tRNAAsp ligase (AMP-forming)
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Antigen T5
-
-
-
-
ASPA
Q52428
gene name
Aspartate--tRNA ligase
-
-
-
-
Aspartic acid translase
-
-
-
-
Aspartic acid translase
-
-
Aspartic acid translase
-
-
Aspartic acid translase
-
-
Aspartic acid translase
-
-
Aspartic acid translase
-
-
Aspartic acid translase
Q52428
-
Aspartic acid translase
Q9LCY8
-
Aspartyl ribonucleate synthetase
-
-
-
-
Aspartyl ribonucleate synthetase
-
-
Aspartyl ribonucleate synthetase
-
-
Aspartyl ribonucleate synthetase
-
-
Aspartyl ribonucleate synthetase
-
-
Aspartyl ribonucleate synthetase
-
-
Aspartyl ribonucleate synthetase
-
-
Aspartyl ribonucleate synthetase
Q52428
-
Aspartyl ribonucleate synthetase
-
-
Aspartyl ribonucleate synthetase
Q9LCY8
-
Aspartyl ribonucleic synthetase
-
-
-
-
aspartyl ribonuleic synthetase
-
-
aspartyl ribonuleic synthetase
-
-
aspartyl ribonuleic synthetase
-
-
aspartyl ribonuleic synthetase
-
-
aspartyl ribonuleic synthetase
-
-
aspartyl ribonuleic synthetase
Q52428
-
aspartyl ribonuleic synthetase
Q9LCY8
-
Aspartyl-transfer ribonucleic acid synthetase
-
-
-
-
Aspartyl-transfer ribonucleic acid synthetase
-
-
Aspartyl-transfer ribonucleic acid synthetase
-
-
Aspartyl-transfer ribonucleic acid synthetase
-
-
Aspartyl-transfer ribonucleic acid synthetase
-
-
Aspartyl-transfer ribonucleic acid synthetase
-
-
Aspartyl-transfer ribonucleic acid synthetase
-
-
Aspartyl-transfer ribonucleic acid synthetase
Q52428
-
Aspartyl-transfer ribonucleic acid synthetase
-
-
Aspartyl-transfer ribonucleic acid synthetase
Q9LCY8
-
Aspartyl-transfer RNA synthetase
-
-
-
-
Aspartyl-transfer RNA synthetase
-
-
Aspartyl-transfer RNA synthetase
-
-
Aspartyl-transfer RNA synthetase
-
-
Aspartyl-transfer RNA synthetase
-
-
Aspartyl-transfer RNA synthetase
-
-
Aspartyl-transfer RNA synthetase
-
-
Aspartyl-transfer RNA synthetase
Q52428
-
Aspartyl-transfer RNA synthetase
-
-
Aspartyl-transfer RNA synthetase
Q9LCY8
-
Aspartyl-tRNA synthetase
-
-
-
-
Aspartyl-tRNA synthetase
-
-
Aspartyl-tRNA synthetase
C4LZN0
-
Aspartyl-tRNA synthetase
Escherichia coli TG1
-
-
-
Aspartyl-tRNA synthetase
-
-
Aspartyl-tRNA synthetase
-
-
Aspartyl-tRNA synthetase
-
-
Aspartyl-tRNA synthetase
P14868
-
Aspartyl-tRNA synthetase
Q6PI48
-
Aspartyl-tRNA synthetase
-
-
Aspartyl-tRNA synthetase
Q3U987
-
Aspartyl-tRNA synthetase
Q8I2B1
-
Aspartyl-tRNA synthetase
-
-
Aspartyl-tRNA synthetase
-
-
Aspartyl-tRNA synthetase
Q52428
-
Aspartyl-tRNA synthetase
Q9LCY8
-
Aspartyl-tRNA synthetase
Q585M3
-
AspRS
-
-
-
-
AspRS
C4LZN0
-
AspRS
P14868
-
AspRS
Q3U987
-
AspRS
Q8I2B1
-
AspRS
Q585M3
-
Bt cyt-AspRS
-
-
cytoplasmic aspartyl-tRNA synthetase
-
-
D-AspRS
-, Q52428
-
discriminating aspartyl-tRNA synthetase
-
-
discriminating aspartyl-tRNA synthetase
Q52428
-
discriminating AspRS
-
-
Ec AspRS
-
-
HS cyt-AspRS
-
-
Hs mt-AspRS
-
-
hsAspRS
P14868
-
mAspRS
-
-
mitochondrial aspartyl-tRNA synthetase
-
-
mitochondrial aspartyl-tRNA synthetase
Q6PI48
-
mmAspRS
Q3U987
-
mt-AspRS
Q6PI48
-
non-discriminating aspartyl-tRNA synthetase
-
-
non-discriminating AspRS
-
-
Synthetase, aspartyl-transfer ribonucleate
-
-
-
-
Tb-AspRS1
Q585M3
-
Tb-AspRS2
Q38CB4
-
TK0492
Q52428
locus name
CAS REGISTRY NUMBER
COMMENTARY
9027-32-1
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
gene aspS
-
-
Manually annotated by BRENDA team
overproducing strain, wild-type and thermosensitive mutant, resulting in substitution of Pro555 by Ser
-
-
Manually annotated by BRENDA team
purified recombinant enzyme from overexpression
-
-
Manually annotated by BRENDA team
strain B
-
-
Manually annotated by BRENDA team
strain TG1
-
-
Manually annotated by BRENDA team
Escherichia coli TG1
strain TG1
-
-
Manually annotated by BRENDA team
expression COS cells
-
-
Manually annotated by BRENDA team
gene DARS2
-
-
Manually annotated by BRENDA team
gene DARS2
UniProt
Manually annotated by BRENDA team
gene DARS2; male, DARS2 gene
-
-
Manually annotated by BRENDA team
wild-type and N-terminal 32-residue truncated form (hDRS delta 32) expressed in Escherichia coli
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
Q3U987
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain KOD1, overexpressed in Escherichia coli
-
-
Manually annotated by BRENDA team
Pyrococcus sp. KOD1
strain KOD1, overexpressed in Escherichia coli
-
-
Manually annotated by BRENDA team
native recombinant AspRS from rat and the N-terminal truncated derivatives AspRS-DELTA20 and AspRS-DELTA36, expressed in yeast
-
-
Manually annotated by BRENDA team
wild-type and mutant enzyme with a deletion of 34 amino acids from its N-terminus
-
-
Manually annotated by BRENDA team
wild-type and two truncated forms lacking 20 or 36 amino acid residues from their amino-terminal polypeptide extension, overexpressed in Saccharomyces cerevisiae
-
-
Manually annotated by BRENDA team
dimeric class II aspartyl-tRNA synthetase
Uniprot
Manually annotated by BRENDA team
dimeric class II aspartyl-tRNA synthetase
-
-
Manually annotated by BRENDA team
mutants with substituted His residues, expressed in Escherichia coli
-
-
Manually annotated by BRENDA team
native and Pro273Gly mutant
-
-
Manually annotated by BRENDA team
native form and truncated form that has lost the first 50-64 residues
-
-
Manually annotated by BRENDA team
recombinant purified enzyme
-
-
Manually annotated by BRENDA team
strain C836
-
-
Manually annotated by BRENDA team
wild-type, C-terminal, and N-terminal truncated forms
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae C836
strain C836
-
-
Manually annotated by BRENDA team
class IIb aminoacyl-tRNA synthetase
-
-
Manually annotated by BRENDA team
isozyme AspRS1
-
-
Manually annotated by BRENDA team
isozyme AspRS2
-
-
Manually annotated by BRENDA team
overexpressed in Escherichia coli; strain HB8; strain VK-1
SwissProt
Manually annotated by BRENDA team
overproduced in Escherichia coli
-
-
Manually annotated by BRENDA team
prokaryotic dimeric class IIb aspartyl-tRNA synthetase
-
-
Manually annotated by BRENDA team
strain HB8
-
-
Manually annotated by BRENDA team
strain HB8, Asp-tRNA synthetase AspRS1
SwissProt
Manually annotated by BRENDA team
strain HB8, Asp-tRNA synthetase AspRS2
SwissProt
Manually annotated by BRENDA team
strain HB8, gene aspS2
-
-
Manually annotated by BRENDA team
Thermus thermophilus VK-1
strain VK-1
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
Q6PI48
LBSL, i.e. leukoencephalopathy with brain stem and spinal cord involvement and lactate elevation, is a monogenic disease associated with a large variety of mutations affecting the human nuclear gene DARS2, encoding mt-AspRS, overview
physiological function
Q6PI48
the mitochondrial aspartyl-tRNA synthetase is a key enzyme for mitochondrial translation and is correlated with leukoencephalopathy
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + Asp + tRNAAsn
AMP + diphosphate + aspartyl-tRNAAsn
show the reaction diagram
-
discriminating AspRS gains the ability to form Asp-tRNAAsn in vitro when the W26H or K85P changes are introduced independently or in combination
-
-
?
ATP + Asp + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + D-aspartate + tRNAAsp
AMP + diphosphate + D-aspartyl-tRNAAsp
show the reaction diagram
-
aspartyl-tRNA synthetase can misacylate tRNAAsp with D-aspartate instead of its usual substrate, L-Asp, substrate specificity and molecular dynamics simulations, overview
-
-
?
ATP + L-Asp + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
low reaction with mutant tRNAAsp with A instead of G at position 73 (tRNAAspA73)
-
-
?
ATP + L-asparagine + tRNAAsp
AMP + diphosphate + L-asparaginyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P36419, Q9LCY8
about half as effective as tRNAAsp
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P36419, Q9LCY8
about have as effective as tRNAAsp
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
the archaeal AspRS enzyme is nondiscriminating, which means that it forms Asp-tRNAAsp and Asp-tRNAAsn, which is the intermediate in AsntRNAAsn generation by ASp-tRNAAsn amidotransferase, in contrary bacterial enzymes are discriminating ones
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
the archaeal AspRS2 enzyme is discriminating, which means that it forms only Asp-tRNAAsp and not Asp-tRNAAsn, the L1 loop exchange mutant is rendered non-dicriminating
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
the archaeal AspRS2 enzyme is nondiscriminating, which means that it forms Asp-tRNAAsp and Asp-tRNAAsn
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
non-discriminating AspRS2, non-discriminating AspRS2, recombinantly produced tRNAAsn
-
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
wild-type enzyme shows no activity with tRNAAsn. Mutant enzymes W26H, K85P and W26H/K85P are active with tRNAAsn
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P36419, -
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P04802
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Q52428
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P36419, Q9LCY8
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Q38CB4, Q585M3, -
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P14868
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-, Q8I2B1
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
C4LZN0, -
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-, Q3U987
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
conformational changes and conformational stability upon tRNA and adenylate binding
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
specificty of tRNA recognition by the enzyme is primarily ensured by the tRNA identity determinants, the discriminator base G37, four bases in the anticodon loop G34, U35, C36, and C38, and G10-U25 base pair in the core region of the tRNA, substrate specificity of wild-type and truncated mutant enzymes, overview
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P36419, Q9LCY8
Thermus thermophilus or Escherichia coli tRNA
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P36419, Q9LCY8
Thermus thermophilus or Eschrichia coli tRNA
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P04802
tRNA undergoes large conformational changes upon binding to the enzyme, specific charging of amino acid resdiue on tRNA, accurate recognition by the enzyme is achieved through sequence and structural signalling
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
wild-type Escherichia coli tRNAAsp and some recombinant acceptor stem mutants from Saccharomyces cerevisiae, overview, protein-RNA recognition between the enzyme and tRNA is highly specific and essential for cell viability
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
specific amino acid binding by the enzyme is required for correct translation of the genetic code
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P04802
the N-terminal extension of each AspRS subunit plays a crucial role in anchoring the tRNA-like motifs of the mRNA on the synthetase
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
aspartyl-tRNA synthetase can misacylate tRNAAsp with D-aspartate instead of its usual substrate, L-Asp, substrate specificity and molecular dynamics simulations, overview
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
the yeast AspRS not only binds and aminoacylates tRNAAsp but also binds its yeast mRNA and initiates retro-inhibition of its expression, overview
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
discriminating AspRS1 and non-discriminating AspRS2
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
the yeast enzyme also binds its own mRNA and autoinhibits itself
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
no activity with tRNAAsn
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
wild-type enzyme shows no activity with tRNAAsn
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Pyrococcus sp. KOD1
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Saccharomyces cerevisiae C836
-
-
-
-
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
enzyme deficiency or mutation is involved in development of autosomal recessive disease leukoencephalopathy with brain stem and spinal cord involvement and lactate elevation, i.e. LBSL, often manifesting in early childhood, overview
-
-
?
GTP + L-aspartate + tRNAAsp
GMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
-
GTP + L-aspartate + tRNAAsp
GMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Q52428
-
-
-
?
UTP + L-aspartate + tRNAAsp
UMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
-
UTP + L-aspartate + tRNAAsp
UMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Q52428
-
-
-
?
UTP + L-aspartate + tRNAAsp
UMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Pyrococcus sp. KOD1
-
-
-
-
-
GTP + L-aspartate + tRNAAsp
GMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Pyrococcus sp. KOD1
-
-
-
-
-
additional information
?
-
-
enzyme can also utilize 2-aminomalonic acid and threo-3-hydroxyaspartic acid in ATP-diphosphate exchange
-
-
-
additional information
?
-
-
aspartate-dependent ATP-diphosphate exchange
-
-
-
additional information
?
-
-
activity of tRNAAsp mimics
-
?
additional information
?
-
-
determination of binding free energies
-
?
additional information
?
-
-
erroneous binding of Asn by the enzyme is highly improbable, determination of binding energy
-
?
additional information
?
-
-
no activity with wild-type tRNAAsp and several acceptor stem mutants from Saccharomyces cerevisiae, overview
-
?
additional information
?
-
-
no acylation with L-aspartate of tRNAAsn
-
?
additional information
?
-
-
aspartyl- and asparaginyl-tRNA synthetases have evolved relatively recently from a comon ancestor
-
?
additional information
?
-
-
no acylation with L-aspartate of tRNAAsn, enzyme is discriminating, because it forms Asn-tRNAAsn by direct aminoacylation, not via the intermediate of Asp-tRNAAsn, and it contains no Asp-tRNAAsn amidotransferase
-
?
additional information
?
-
-
the enzyme also binds to its mRNA via its N-terminal extension of 70 amino acid residues and the anticodon-binding module, which has a regulatory function on the expression of the enzyme
-
?
additional information
?
-
-
the N-terminal extension of the enzyme is involved in the transfer of Asp-tRNAAsp to elongation factor alpha1, the structural switch model supports the direct transfer mechanism
-
?
additional information
?
-
-
no transfer of lysine on tRNAAsp
-
-
-
additional information
?
-
-
in vitro transcription-translation activity and inhibition by Microcystin C, overview
-
-
-
additional information
?
-
P21889
the co-substrate ATP binds preferentially with three associated Mg2+ cations in an unusual, bent geometry, the Mg2+ cations play a structural role and also participate catalytically in the enzyme reaction, co-binding of the ATP-Mg3+ complex increases the Asp/Asn binding free energy difference, indicating that amino acid discrimination is substrate-assisted, molecular dynamics simulations, overview
-
-
-
additional information
?
-
-
AspRS mediates stimulation of lysyl-tRNA synthetase, KRShe, with 40% stimulation when eight fold excess of AspRS is present. The non-synthetase protein from the multi-synthetase complex p38 inhibits the AspRS-mediated stimulation
-
-
-
additional information
?
-
-
in Thermus thermophilus Asn-tRNAAsn is formed indirectly via a two-step pathway whereby tRNAAsn is mischarged with Asp that will subsequently be amidated into Asn by an amidotransferase.The non-discriminating aspartyl-tRNA synthetase, the trimeric GatCAB tRNA-dependent amidotransferase and the tRNAAsn promoting this pathway assemble into a ribonucleoprotein particle termed transamidosome, analysis of the mechanism of Asn-tRNAAsn formation by the transamidosome, overview
-
-
-
additional information
?
-
-
misaspartylation of tRNAAsn and tRNAGlu does not exist in vivo
-
-
-
additional information
?
-
-
mutations in the DARS2 gene cause the autosomal recessive disorder leukoencephalopathy with brainstem and spinal cord involvement and lactate elevation, overview
-
-
-
additional information
?
-
-
ATP-diphosphate exchange activity of wild-type and mutant enzymes in the presence of Asp or Asn, overview
-
-
-
additional information
?
-
-
increased concentrations of AspRS lead to the accumulation of significant amounts of Asp-tRNAAsn and Asp-tRNAGlu in vitro, but not in vivo. The enzyme does not perform autoaspartylation
-
-
-
additional information
?
-
-
Thermus thermophilus contains two AspRSs: the discriminating AspRS1 which aspartylates only tRNAAsp and the non-discriminating AspRS2 which aspartylates tRNAAsn as efficiently as tRNAAsp
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
the archaeal AspRS enzyme is nondiscriminating, which means that it forms Asp-tRNAAsp and Asp-tRNAAsn, which is the intermediate in AsntRNAAsn generation by ASp-tRNAAsn amidotransferase, in contrary bacterial enzymes are discriminating ones
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
the archaeal AspRS2 enzyme is discriminating, which means that it forms only Asp-tRNAAsp and not Asp-tRNAAsn, the L1 loop exchange mutant is rendered non-dicriminating
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
the archaeal AspRS2 enzyme is nondiscriminating, which means that it forms Asp-tRNAAsp and Asp-tRNAAsn
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
non-discriminating AspRS2
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P04802
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P36419, Q9LCY8
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
Q38CB4, Q585M3, -
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
P14868
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-, Q8I2B1
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
C4LZN0, -
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-, Q3U987
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
protein-RNA recognition between the enzyme and tRNA is highly specific and essential for cell viability
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
specific amino acid binding by the enzyme is required for correct translation of the genetic code
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
the yeast AspRS not only binds and aminoacylates tRNAAsp but also binds its yeast mRNA and initiates retro-inhibition of its expression, overview
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
-
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
enzyme deficiency or mutation is involved in development of autosomal recessive disease leukoencephalopathy with brain stem and spinal cord involvement and lactate elevation, i.e. LBSL, often manifesting in early childhood, overview
-
-
?
additional information
?
-
-
aspartyl- and asparaginyl-tRNA synthetases have evolved relatively recently from a comon ancestor
-
?
additional information
?
-
-
no acylation with L-aspartate of tRNAAsn, enzyme is discriminating, because it forms Asn-tRNAAsn by direct aminoacylation, not via the intermediate of Asp-tRNAAsn, and it contains no Asp-tRNAAsn amidotransferase
-
?
additional information
?
-
-
the enzyme also binds to its mRNA via its N-terminal extension of 70 amino acid residues and the anticodon-binding module, which has a regulatory function on the expression of the enzyme
-
?
additional information
?
-
-
the N-terminal extension of the enzyme is involved in the transfer of Asp-tRNAAsp to elongation factor alpha1, the structural switch model supports the direct transfer mechanism
-
?
additional information
?
-
-
AspRS mediates stimulation of lysyl-tRNA synthetase, KRShe, with 40% stimulation when eight fold excess of AspRS is present. The non-synthetase protein from the multi-synthetase complex p38 inhibits the AspRS-mediated stimulation
-
-
-
additional information
?
-
-
in Thermus thermophilus Asn-tRNAAsn is formed indirectly via a two-step pathway whereby tRNAAsn is mischarged with Asp that will subsequently be amidated into Asn by an amidotransferase.The non-discriminating aspartyl-tRNA synthetase, the trimeric GatCAB tRNA-dependent amidotransferase and the tRNAAsn promoting this pathway assemble into a ribonucleoprotein particle termed transamidosome, analysis of the mechanism of Asn-tRNAAsn formation by the transamidosome, overview
-
-
-
additional information
?
-
-
misaspartylation of tRNAAsn and tRNAGlu does not exist in vivo
-
-
-
additional information
?
-
-
mutations in the DARS2 gene cause the autosomal recessive disorder leukoencephalopathy with brainstem and spinal cord involvement and lactate elevation, overview
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
the co-substrate ATP binds preferentially with three associated Mg2+ cations in an unusual, bent geometry, co-binding of the ATP-Mg3+ complex increases the Asp/Asn binding free energy difference
ATP
Q38CB4, Q585M3, -
;
ATP
-, Q3U987
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
-
can replace Mg2+, with 12% efficiency in activation of ATP-diphosphate exchange, with 34% efficiency in aminoacylation
K2HPO4
-
enzyme form PC-3 is stimulated, optimal concentration 75 mM, enzyme form PC-1 and enzyme form PC-2 are inhibited
KCl
-
enzyme form PC-3 is stimulated, optimal concentration 75 mM, enzyme form PC-1 and enzyme form PC-2 are inhibited
Mg2+
-
Km: 9 mM; required
Mg2+
-
Km: 1.1 mM , aminoacylation; Km: 6.9 mM , ATP-diphosphate exchange; optimal concentration: 12 mM, ATP-diphosphate exchange, 5 mM, aminoacylation; required
Mg2+
-
optimal Mg2+/ATP ratio is 2; required
Mg2+
-
the co-substrate ATP binds preferentially with three associated Mg2+ cations in an unusual, bent geometry, the Mg2+ cations play a structural role and also participate catalytically in the enzyme reaction, co-binding of the ATP-Mg3+ complex increases the Asp/Asn binding free energy difference
Mg2+
Q6PI48
required
Mn2+
-
can replace Mg2+, with 35% efficiency in activation of ATP-diphosphate exchange, with 56% efficiency in aminoacylation
PO43-
-
appears to act synergistically with K+ in stimulation of enzyme form PC-3, no effect on either enzyme form PC-1 or enzyme form PC-2
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(3S)-3-amino-5-((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)oxy)(hydroxy)phosphoryl)-4-oxopentanoic acid
-
-
2-aminomalonic acid
-
inhibits transfer of Asp to tRNA
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
-
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
ASP-AMS
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
ASP-AMS
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-, Q8I2B1
ASP-AMS
Br-
-
0.7 M, 50% inhibition of ATP-diphosphate exchange
CaCl2
-
1 mM, more than 50% inhibition
CH3COO-
-
2.0 M, 50% inhibition of ATP-diphosphate exchange
Cl-
-
2.2 M, 50% inhibition of ATP-diphosphate exchange
ClO3-
-
0.1 M, 50% inhibition of ATP-diphosphate exchange
CNS-
-
0.1 M, 50% inhibition of ATP-diphosphate exchange
Cs+
-
1.1 M, 50% inhibition of ATP-diphosphate exchange, 0.16 M, 50% inhibition of aminoacylation
diethyldicarbonate
-
reversed by hydroxylamine
erythro-3-hydroxyaspartic acid
-
inhibits transfer of Asp to tRNA
heptapeptide-nucleotide microcin C
-
i.e. McC, upon its entry into a susceptible cell, McC is processed to release a nonhydrolyzable aspartyl-adenylate that inhibits aspartyl-tRNA synthetase, leading to the cessation of translation and cell growth. The rate-limiting step of McC processing in vitro is deformylation of the first methionine residue of McC
I-
-
0.1 M, 50% inhibition of ATP-diphosphate exchange
IO3-
-
0.04 M, 50% inhibition of ATP-diphosphate exchange
K+
-
2.3 M, 50% inhibition of ATP-diphosphate exchange, 0.22 M, 50% inhibition of aminoacylation
K2HPO4
-
inhibits enzyme forms PC-1 and PC-2, PC-3 is stimulated (optimal concentration 75 mM)
KCl
-
inhibits enzyme forms PC-1 and PC-2, PC-3 is stimulated (optimal concentration 75 mM)
L-aspartol adenylate
-
-
L-aspartol-adenylate
-
aspartol-AMP
L-aspartol-adenylate
-
aspartol-AMP
L-aspartol-adenylate
-, Q8I2B1
aspartol-AMP
L-aspartyl adenylate
-
-
Li+
-
0.6 M, 50% inhibition of ATP-diphosphate exchange, 0.11 M, 50% inhibition of aminoacylation
Microcin C
-
a ribosome-synthesized heptapeptide that contains a modified adenosine monophosphate covalently attached to the C-terminal aspartate, a potent inhibitor of bacterial growth, that targets the translation process via its degradation product, a modified aspartyl-adenylate containing an N-acylphosphoramidate linkage, overview, Microcystin is recombinantly expressed in Escherichia coli strain TG1
-
Na+
-
1.4 M, 50% inhibition of ATP-diphosphate exchange, 0.18 M , 50% inhibition of aminoacylation
NaCl
-
inhibits enzyme forms PC-1 and PC-2 much more than PC-3
NH4+
-
0.9 M, 50% inhibition of ATP-diphosphate exchange, 0.14 M, 50% inhibition of aminoacylation
NiCl2
-
0.1 mM more than 50% inhibition
NO3-
-
0.7 M, 50% inhibition of ATP-diphosphate exchange
p-chloromercuribenzoate
-
-
RNAi
Q38CB4, Q585M3, -
;
-
succinate
-
50% inhibition at 210 mM
threo-3-hydroxyaspartic acid
-
inhibits transfer of Asp to tRNA
threo-3-Methylaspartic acid
-
inhibits transfer of Asp to tRNA
tRNA
-
above 0.4 mg/ml
microcin C7-C51
-
an antimicrobial nucleotide peptide that targets aspartyl-tRNA synthetase and inhibits translation. Fragmentation and analysis of fragment structure and inhibitory potency, overview
-
additional information
-
the yeast AspRS initiates retro-inhibition of its expression, overview
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
putrescine
-
enhances activity, optimum concentrations is 0.07 mM
-
putrescine
-
can replace Mg2+ in activation with 27% efficiency in aminoacylation
-
spermidine
-
enhances activity, optimum concentrations is 0.07 mM
spermidine
-
can replace Mg2+ in activation with 16% efficiency in aminoacylation
spermine
-
enhances activity, optimum concentrations is 0.005 mM
spermine
-
can replace Mg2+ in activation with 32% efficiency in aminoacylation
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
8
-
2-aminomalonic acid
-
ATP-diphosphate exchange
0.003
-
Asp
-
37C and 70C, tRNA aminoacylation, enzyme form AspRS2
0.009
-
Asp
-
37C, tRNA aminoacylation, enzyme form AspRS1
0.029
-
Asp
-
truncated enzyme
0.03
-
Asp
-
37C, ATP-diphosphate exchange, and 70C, tRNAaminoacylation, enzyme form AspRS1
0.05
-
Asp
-
-
0.06
-
Asp
-
wild-type enzyme
0.062
-
Asp
-
37C, ATP-diphosphate exchange, enzyme form AspRS2
0.067
-
Asp
-
-
0.1
-
Asp
-
70C, ATP-diphosphate exchange by enzyme form AspRS1, and tRNA aminoacylation by enzyme form AspRS2
0.302
-
Asp
-
native enzyme
0.4
-
Asp
-
wild-type
4.6
-
Asp
-
ATP-diphosphate exchange
1.2
-
aspartate
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme E118K
1.3
-
aspartate
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme A471T; pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), wild-type enzyme
1.4
-
aspartate
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme E93K; pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme L30F
0.03
-
aspartic acid
P36419, Q9LCY8
pH 7.2, 70C
0.033
-
ATP
P36419, Q9LCY8
pH 7.2, 70C
0.04
-
ATP
-
wild-type enzyme
0.05
-
ATP
-
-
0.05
-
ATP
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme E93K
0.09
-
ATP
-
wild-type enzyme
0.091
-
ATP
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), wild-type enzyme
0.092
-
ATP
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme L30F
0.1
-
ATP
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme E118K
0.12
-
ATP
-
37C, tRNA aminoacylation, enzyme form AspRS1
0.14
-
ATP
-, Q8I2B1
Plasmodium falciparum wild type aspartyl-tRNA synthetase, amino acids 49-626
0.25
-
ATP
-
70C, ATP-diphosphate exchange, enzyme form AspRS1
0.26
-
ATP
-
-
0.28
-
ATP
-
70C, tRNA aminoacylation, enzyme form AspRS1
0.28
-
ATP
P36419, Q9LCY8
pH 7.2, 70C
0.37
-
ATP
-
37C, ATP-diphosphate exchange, enzyme form AspRS1
0.515
-
ATP
-
Homo sapiens wild type aspartyl-tRNA synthetase, amino acids 1-500
1.11
-
ATP
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme A471T
0.32
-
L-Asp
-
-
0.0015
-
L-aspartate
-
-
0.016
-
L-aspartate
-
Homo sapiens wild type aspartyl-tRNA synthetase, amino acids 1-500
0.024
-
L-aspartate
-
-
0.037
-
L-aspartate
-
-
0.09
-
L-aspartate
-
-
0.1
-
L-aspartate
-
-
0.21
-
L-aspartate
-, Q8I2B1
Plasmodium falciparum wild type aspartyl-tRNA synthetase, amino acids 49-626
24
-
threo-3-hydroxyaspartic acid
-
ATP-diphosphate exchange
6.3e-05
-
tRNAAsn
P36419, Q9LCY8
pH 7.2, 70C
0.0002
-
tRNAAsn
-
recombinant wild-type enzyme, pH 7.2, 37C
0.0009
-
tRNAAsn
-
recombinant mutant enzyme, pH 7.2, 37C
0.0031
-
tRNAAsn
-
pH 6.0, 60C, mutant enzyme K85P; pH 6.0, 60C, mutant enzyme W26H/K85P
0.0034
-
tRNAAsn
P36419, Q9LCY8
pH 7.2, 70C
0.0062
-
tRNAAsn
-
pH 6.0, 60C, mutant enzyme W26H/K85P; pH 6.0, 60C, mutant enzyme W26HK85P
0.0091
-
tRNAAsn
-
pH 6.0, 60C, mutant enzyme K85P; pH 6.0, 60C, mutant enzyme W26H
1.3e-05
-
tRNAAsp
-
native enzyme
1.8e-05
-
tRNAAsp
-, Q8I2B1
Plasmodium falciparum wild type aspartyl-tRNA synthetase, amino acids 49-626
3e-05
-
tRNAAsp
P36419, Q9LCY8
pH 7.2, 70C
6.1e-05
-
tRNAAsp
-
-
7.3e-05
-
tRNAAsp
P36419, Q9LCY8
pH 7.2, 70C
0.00013
-
tRNAAsp
-
truncated enzyme
0.00013
-
tRNAAsp
-
human mitochondrial tRNAAsp
0.00014
-
tRNAAsp
-
recombinant wild-type enzyme, pH 7.2, 37C
0.00015
-
tRNAAsp
-
Escherichia coli native tRNAAsp
0.00016
-
tRNAAsp
-
Homo sapiens wild type aspartyl-tRNA synthetase, amino acids 1-500
0.0002
-
tRNAAsp
-
recombinant wild-type enzyme, pH 7.2, 37C
0.00027
-
tRNAAsp
-
-
0.0004
-
tRNAAsp
-
-
0.0005
-
tRNAAsp
-
aspartyl-tRNA synthetase as a component of the multisynthetase complex or recombinant native enzyme
0.0006
-
tRNAAsp
-
tRNAAsp, , truncated enzymes AspRS-DELTAN36 and AspRS-DELTAN20
0.0006
-
tRNAAsp
-
wild-type enzyme
0.00072
-
tRNAAsp
-
pH 7.5, 37C, recombinant wild-type enzyme
0.00074
-
tRNAAsp
-
pH 7.5, 37C, recombinant mutant C152F
0.001
-
tRNAAsp
-
recombinant mutant enzyme, pH 7.2, 37C
0.0012
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme W26HK85P; pH 6.0, 60C, wild-type enzyme
0.00128
-
tRNAAsp
-
pH 7.5, 37C, recombinant mutant Q184K
0.0014
-
tRNAAsp
-
pH 6.0, 60C, wild-type enzyme; pH 6.0, 60C, wild-type enzyme
0.0024
-
tRNAAsp
-, Q8I2B1
Plasmodium falciparum aspartyl-tRNA synthetase, mutated RNA binding motif
0.0045
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme W26H; pH 6.0, 60C, mutant enzyme W26H
0.011
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme K85P; pH 6.0, 60C, mutant enzyme K85P
0.08
-
tRNAAsp
-
pH 7.8, 28C
1.3
-
tRNAAspA73
-
pH 7.8, 28C
-
0.005
-
L-aspartic acid
P36419, Q9LCY8
pH 7.2, 70C
additional information
-
additional information
-
Km value of pure tRNA, transcript tRNA and unfractionated tRNAs of various origins with enzyme form AspRS1 and enzyme form AspRS2
-
additional information
-
additional information
-
Km values of ATP-diphosphate exchange reaction and acylation of homodimers and heterodimers of AspRS molecules
-
additional information
-
additional information
-
kinetics, tRNAAsp mimics
-
additional information
-
additional information
-
kinetics of several tRNAAsp variants
-
additional information
-
additional information
-
very low kinetic effiency at 17C
-
additional information
-
additional information
-
ligand binding thermodynamics and kinetics, overview
-
additional information
-
additional information
-
kinetics, thermodynamics of wild-type enzyme and mutant Q199E
-
additional information
-
additional information
-
ligand binding, kinetics and thermodynamics, wild-type and mutant enzymes, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2
8
Asp
-
mutant enzyme
18
-
Asp
-
wild-type enzyme
16.9
-
aspartyl-tRNA
-
-
-
5.9
-
ATP
-, Q8I2B1
Plasmodium falciparum wild type aspartyl-tRNA synthetase, amino acids 49-626, ATP-diphosphate exchange
6.7
-
ATP
-
Homo sapiens wild type aspartyl-tRNA synthetase, amino acids 1-500, ATP-diphosphate exchange
0.0008
-
tRNAAsn
-
recombinant mutant enzyme, pH 7.2, 37C
0.0056
-
tRNAAsn
-
recombinant wild-type enzyme, pH 7.2, 37C
0.009
-
tRNAAsn
-
pH 6.0, 60C, mutant enzyme K85P; pH 6.0, 60C, mutant enzyme W26H/K85P
0.026
-
tRNAAsn
-
pH 6.0, 60C, mutant enzyme K85P; pH 6.0, 60C, mutant enzyme W26H
0.037
-
tRNAAsn
-
pH 6.0, 60C, mutant enzyme W26H/K85P; pH 6.0, 60C, mutant enzyme W26HK85P
0.092
-
tRNAAsn
P36419, Q9LCY8
pH 7.2, 70C
0.12
-
tRNAAsn
P36419, Q9LCY8
pH 7.2, 70C
0.00016
-
tRNAAsp
-
pH 7.5, 37C, recombinant mutant Q184K
0.0016
-
tRNAAsp
-
pH 7.5, 37C, recombinant mutant C152F
0.0083
-
tRNAAsp
-
recombinant mutant enzyme, pH 7.2, 37C
0.01
-
tRNAAsp
-, Q8I2B1
Plasmodium falciparum aspartyl-tRNA synthetase, mutated RNA binding motif
0.0117
-
tRNAAsp
-
recombinant wild-type enzyme, pH 7.2, 37C
0.027
-
tRNAAsp
-
pH 7.5, 37C, recombinant wild-type enzyme
0.034
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme W26H/K85P; pH 6.0, 60C, mutant enzyme W26HK85P
0.042
-
tRNAAsp
-
recombinant wild-type enzyme, pH 7.2, 37C
0.05
-
tRNAAsp
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme E93K
0.061
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme K85P; pH 6.0, 60C, mutant enzyme K85P; pH 6.0, 60C, wild-type enzyme; pH 6.0, 60C, wild-type enzyme
0.1
-
tRNAAsp
-
human mitochondrial tRNAAsp
0.116
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme W26H; pH 6.0, 60C, mutant enzyme W26H
0.133
-
tRNAAsp
-
Escherichia coli native tRNAAsp
0.2
-
tRNAAsp
-, Q8I2B1
Plasmodium falciparum wild type aspartyl-tRNA synthetase, amino acids 49-626
0.22
-
tRNAAsp
-
Homo sapiens wild type aspartyl-tRNA synthetase, amino acids 1-500
0.24
-
tRNAAsp
P36419, Q9LCY8
pH 7.2, 70C
0.55
-
tRNAAsp
-
pH 7.8, 28C
2.7
-
tRNAAsp
P36419, Q9LCY8
pH 7.2, 70C
8
-
tRNAAsp
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme A471T
9.3
-
tRNAAsp
-
wild-type enzyme, acylation activity, pH 7.5, 37C
12
-
tRNAAsp
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme L30F; pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), wild-type enzyme
79
-
tRNAAsp
-
pH 7.5, 37C, aminoacylation of tRNAAsp(QUC), mutant enzyme E118K
0.0029
-
tRNAAspA73
-
pH 7.8, 28C
-
8.5
-
ATP
-, Q8I2B1
Plasmodium falciparum aspartyl-tRNA synthetase, mutated RNA binding motif, ATP-diphosphate exchange
additional information
-
additional information
-
-
-
additional information
-
additional information
-
-
-
additional information
-
additional information
-
activity of tRNAAsp mimics
-
additional information
-
additional information
-
catalytic efficiency of several tRNAAsp variants
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2.9
-
tRNAAsn
-
pH 6.0, 60C, mutant enzyme K85P; pH 6.0, 60C, mutant enzyme W26H
17502
6
-
tRNAAsn
-
pH 6.0, 60C, mutant enzyme W26HK85P
17502
5.8
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme K85P
17503
26
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme W26H
17503
28
-
tRNAAsp
-
pH 6.0, 60C, mutant enzyme W26HK85P
17503
45
-
tRNAAsp
-
pH 6.0, 60C, wild-type enzyme
17503
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.000123
-
(3S)-3-amino-5-((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)oxy)(hydroxy)phosphoryl)-4-oxopentanoic acid
-
-
9.8e-06
-
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
-
1.5e-05
-
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
pH 7.5, 37C
1.5e-05
-
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
-
7.9e-05
-
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
Homo sapiens wild type aspartyl-tRNA synthetase, amino acids 1-500
0.00026
-
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-, Q8I2B1
Plasmodium falciparum wild type aspartyl-tRNA synthetase, amino acids 49-626
0.00028
-
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
-
0.00039
-
5'-O-[N-(L-aspartyl)sulfamoyl]adenosine
-
-
1
-
CaCl2
-
-
0.045
-
L-aspartol adenylate
-
pH 7.5, 37C
0.00033
-
L-aspartol-adenylate
-
Homo sapiens wild type aspartyl-tRNA synthetase, amino acids 1-500
0.0008
-
L-aspartol-adenylate
-, Q8I2B1
Plasmodium falciparum wild type aspartyl-tRNA synthetase, amino acids 49-626
0.0046
-
L-aspartol-adenylate
-
-
0.01
-
L-aspartol-adenylate
-
-
0.027
-
L-aspartol-adenylate
-
-
0.041
-
L-aspartol-adenylate
-
-
0.045
-
L-aspartol-adenylate
-
-
0.1
-
NiCl2
-
-
6000
-
Li+
-
ATP-diphosphate exchange
additional information
-
additional information
-
Microcystin C in vivo inhibition kinetics
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2e-05
-
P36419, Q9LCY8
native enzyme, cell extract, 37C
8.3e-05
-
P36419, Q9LCY8
native enzyme, cell extract, 37C
0.017
-
-
wild-type enzyme
0.025
-
-
recombinant wild-type enzyme
0.0336
-
P36419, Q9LCY8
purified recombinant enzyme, 37C
0.034
-
-
truncated enzyme
0.0405
-
-
enzyme form AspRS2
0.043
-
P36419, Q9LCY8
recombinant enzyme, Escherichia coli DH5alpha cell extract, 37C
0.0452
-
-
enzyme form AspRS1
0.055
-
P36419, Q9LCY8
recombinant enzyme, Escherichia coli BL21 cell extract, 37C
0.063
-
-
recombinant mutant D560V
0.097
-
-
recombinant mutant R263Q
0.11
-
-
65C, pH is not specified in the publication; 65C, pH not specified in the publication
0.12
-
-
recombinant mutant L626V
0.124
-
P36419, Q9LCY8
recombinant enzyme, Escherichia coli BL21 cell extract, 37C
0.15
-
-
recombinant mutant Q184K
0.155
-
P36419, Q9LCY8
recombinant enzyme, Escherichia coli DH5alpha cell extract, 37C
0.321
-
P36419, Q9LCY8
purified recombinant enzyme, 37C
0.395
-
-
-
1.75
-
-
recombinant mutant C152F
3.7
-
-
purified recombinant His6-tagged DRS enzyme
10.9
-
-
purified recombinant His6-tagged DRS-ubiquitin fusion enzyme
11.5
-
-
purified recombinant biotin-tagged DRS-ubiquitin fusion enzyme
13.5
-
-
purified recombinant His6-tagged DRS-small ubiquitin-like modifier fusion enzyme
17
-
-
purified recombinant GST-tagged DRS enzyme
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
activity of tRNAAsp mimics
additional information
-
-
-
additional information
-
-
activity with several tRNAAsp variants
additional information
-
-
ATP-diphosphate exchange activity of wild-type and mutant enzymes in the presence of Asp or Asn, overview
additional information
-
Q38CB4, Q585M3, -
100% aminoacylation of cytosolic tRNA, 4% aminoacylation of mitochondrial tRNA; 100% aminoacylation of cytosolic tRNA, 90% aminoacylation of mitochondrial tRNA
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.2
-
P36419, Q9LCY8
assay at
7.2
-
-
assay at
7.3
-
Q6PI48
assay at
7.4
-
-
assay at
7.4
-
-
ATP/diphosphate exchange and tRNA aminoacylation assay
7.4
-
Q8I2B1
ATP/diphosphate exchange and tRNA aminoacylation assay
7.5
-
-
assay at
7.5
-
-
assay at
7.5
-
-
aspartylation and inhibition assay
7.6
-
-
assay at
7.8
8
-
potassium Hepes buffer
8
-
-
ATP-diphosphate exchange
8
-
-
assay at
8.5
-
-
Tris/cacodylate buffer
additional information
-
-
very low kinetic effiency at 17C
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
8
-
pH 5.5: about 60% of maximal activity, pH 8.0: about 65% of maximal activity
5.5
9
-
pH 5.5: about 60% of maximal activity, pH 9.0: about 30% of maximal activity
7.8
9.1
-
7.8: sharp loss of activity below, 9.1: plateau of high activity up to pH 9.1
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
25
-
-
assay at
30
-
-
assay at
30
-
Q6PI48
assay at
37
50
-
assay at 37C or 50C
37
-
-
assay at
37
-
-
aspartylation and inhibition assay
37
-
-
ATP/diphosphate exchange and tRNA aminoacylation assay
37
-
Q8I2B1
ATP/diphosphate exchange and tRNA aminoacylation assay
65
-
-
-
65
-
-
optimum temperature shifts from 65C to 75C in the presence of polyamine molecules putrescine, spermine or spermidine
75
-
-
in presence of polyamine molecules, such as putrescine, spermine, and spermidine
75
-
-
optimum temperature shifts from 65C to 75C in the presence of polyamine molecules putrescine, spermine or spermidine
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
75
-
35C: about 60% of maximal activity, 75C: about 50% of maximal activity
45
85
-
45C: about 80% of maximal activity, 85C: about 25% of maximal activity
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.23
-
-
calculated from sequence
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Sulfolobus tokodaii (strain DSM 16993 / JCM 10545 / NBRC 100140 / 7)
Thermococcus kodakaraensis (strain ATCC BAA-918 / JCM 12380 / KOD1)
Thermococcus kodakaraensis (strain ATCC BAA-918 / JCM 12380 / KOD1)
Thermococcus kodakaraensis (strain ATCC BAA-918 / JCM 12380 / KOD1)
Thermococcus kodakaraensis (strain ATCC BAA-918 / JCM 12380 / KOD1)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
50000
-
-
determined by SDS-PAGE
60800
-
Q38CB4, Q585M3, -
theoretical
62900
-
Q38CB4, Q585M3, -
theoretical
64000
-
-
determined by SDS-PAGE
66000
-
-, Q8I2B1
determined by SDS-PAGE
83000
100000
-
sucrose density gradient centrifugation
89000
106000
-
sucrose density gradient centrifugation
96000
-
-
crystal structure determination
116000
-
-
sedimentation equilibrium determination, neutron scattering
117000
-
-
gel filtration, also a smaller peak of MW 57000 detected
119000
-
-
sedimentation equilibrium determination
122000
-
-
nondenaturing PAGE
122000
-
-
crystal structure determination
124000
-
-
gel filtration
132000
-
-
gel filtration
150000
-
-
gel filtration, PC-1 and PC-2
156000
-
-
nondenaturing PAGE
205000
-
-
gel filtration
260000
-
-
gel filtration
500000
-
-
gel filtration, PC-3, possibly PC-3 represents a large complex of aminoacyl-tRNA synthetases
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
P36419
x * 66030, calculation from nucleotide sequence
?
P36419, Q9LCY8
x * 48329, amino acid sequence determination; x * 66029, amino acid sequence determination
?
-
x * 65912, crystal structure determination
?
-
x * 50000, calculated from sequence; x * 50000, SDS-PAGE; x * 50893, calculated from sequence
dimer
-
2 * 50000, SDS-PAGE
dimer
-
2 * 61000, sedimentation equilibrium under denaturing conditions; 2 * 64000, PAGE under denaturing conditions
dimer
-
2 * 53000, gel electrophoresis under denaturing conditions
dimer
-
2 * 63000, SDS-PAGE
dimer
-
homodimer
dimer
-
modular structure
dimer
-, Q8I2B1
-
homodimer
C4LZN0, -
-
monomer
-
1 * 106000-114000, SDS-PAGE
monomer
Saccharomyces cerevisiae C836
-
1 * 106000-114000, SDS-PAGE
-
additional information
-
enzymes exist as free soluble enzymes
additional information
-
aspartyl-tRNA synthetase is a component of a multienzyme complex comprising nine aminoacyl-tRNA synthetases
additional information
-
enzyme occurs in the multienzyme complex of aminoacyl-tRNA synthetases
additional information
-
enzyme is part of a high molecular weight aminoacyl-tRNA synthetase complex, which is resistant to dissociation when subjected to gel filtration
additional information
-
enzyme is part of a high molecular mass aminoacyl-tRNA synthetase complex, which has a coherrent structure, that can be visualized by electron microscopy
additional information
P36419, Q9LCY8
modular organization
additional information
-
secondary structure determination, structural switch model, the C-terminus adopts a regular alpha-helix with amphiphilicity, while the N-terminus shows a less-ordered structure with a flexible beta turn
additional information
-
the N-terminal appendix of 70 amino acid residues adopts a helical structure and encompasses the RNA binding motif XSKXXLKKXK
additional information
-
domain structure
additional information
-
the N-terminal 70 amino acid residues extension protrudes from the anticodon-binding module, it is not essential for acylation activity but contains the RNA-binding motif that promotes non-specific interactions with the tRNAs, it can also interact with the 5'-end of the enzyme's mRNA
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
no glycoprotein
-
-
lipoprotein
-
high molecular weight aminoacyl-tRNA synthetase complex contains lipid. Delipidation does not affect the size or activity of the complex, but a variety of functional and structural properties of individual synthetases in the complex are altered: sensitivity to salts plus detergents, temperature inactivation, hydrophobicity, sensitivity to protease digestion
additional information
-
the enzyme does not perform autoaspartylation in vivo
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the crystal structure of aspartyl-tRNA synthetase is determined at 2.8 A resolution
C4LZN0, -
analysis of the 2.6-A resolution crystal structure of Escherichia coli AspRS with bound aspartyl-adenylate, AspAMP, molecular dynamics simulations
-
of enzyme complexed with its cognate tRNA
-
purified recombinant enzyme, in ammonium sulfate, NaCl, bis Tris propane, isopropanol, X-ray diffraction structure determination at 2.7 A resolution and structure analysis
-
binary complex formed by the enzyme and tRNAAsp
-
purified recombinant truncated enzyme, tri- and tetragonal crystals, X-ry structure determination at 3 and 2.3 A resolution, respectively, molecular replacement, structure analysis and modeling
P04802
7 different mutants, modification of crystal surfaces, investigation of crystallizability to determine the influence of surface residues and protein structure on crystal growth, packing arrangement, and quality
-
AspRS2, hanging drop vapour diffusion method, 0.002 ml of 14 mg/ml protein with 10% m/v PEG 8000, 200 mM NaCl, and 100 mM Na-CHES buffer, pH 9.5, X-ray diffraction structure determination and analysis at 2.3-3.2 A resolution
-
at 2.5 A resolution
-
enzyme complexed with a non-hydrolysable analogue of asparaginyl-adenylate and with ATP, X-ray diffraction structure determination at 2.6 A resolution
-
hanging-drop vapour-diffusion method, enzyme crystallizes either in a monoclinic or an orthorhombic habit. Minute amounts of protein impurities alter to a different extent the growth of each crystal form. The best synthetase crystals are obtained when the crystallizating solution is either enclosed in capillaries or immobilized in agarose gels
-
isozyme AspRS2, hanging-drop vapour diffusion method, pH 9.5, in presence of PEG 8000 and NaCl, structure determination
-
purified enzyme complexed with tRNAAsp from Thermus thermophilus or Escherichia coli, potential intermediate of the recognition process, protein solution: 15 mg/ml of enzyme, 7.35 mg/ml of tRNA, plus equal volume of reservoir solution: 10 mM MgCl2, 50 mM HEPES, pH 7.5, 0.7 M sodium citrate, 17C, 2 weeks, X-ray diffraction structure determination at 3.5 A resolution, structure analysis
-
purified recombinant enzyme, hanging-drop vapour diffusion method, in presence of PEG 8000, at 293K, growth kinetics and solubility measurements, X-ray diffraction structure determination at 3.1 A resolution
-
purified wild-type and seleno-Met isozymes AspRS2, vapour phase diffusion from mother liquid: 100 mM CHES buffer, pH 9.5, 200 mM NaCl, 10% w/v PEG 8000, X-ray diffraction structure determination at 2.3 A resolution, structure analysis
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
native enzyme is stable for 24 h, half-life of truncated enzyme hDRSDELTA32 is 7 h, half-life of the fusion protein of glutathione-S-transferase and hDRSDELTA32 is 3 h
42
-
-
half-life: 22 min for tRNA aminoacylation, 68 min for ATP/diphosphate exchange
50
-
-
half-life: 7 min without stabilizing reagents, 12 min in presence of 0.004 mM ATP, 23 min in presence of 0.004 mM ATP and 0.1 mM Asp
55
-
-
10 min stable, 95% loss of activity after 2 h
70
-
-
no denaturation below
85
-
-
the thermal melting temperature seems higher than 85C in the presence of Mg2+. In the absence of Mg 2+, melting temperature decreases to 85C
90
-
-
half-life is 17 h
90
-
-
half-life: 17 h
100
-
-
half-life is 3.5 h
100
-
-
half-life: 3.5 h
additional information
-
-
thermostable isozyme AspRS1
additional information
-
-
thermosensitive mutant, resulting in substitution of Pro 555 by Ser
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
polyamines stabilize the conformation of the tRNA-AspRS complex
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Glycerol
-
25% v/v, required throughout purification procedure to maintain stability
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80C, 0.1 M potassium phosphate buffer, about 50% loss of activity after 6 months
-
-20C, 10 mM potassium phosphate, pH 7.6, 10 mM 2-mercaptoethanol, 50% glycerol, stable
-
-20C, 40% w/v glycerol, SH-group stabilizing reagent, 50% loss of activity after 72 h
-
in a dry form, e.g. as acetone precipitate, stable for 1 month
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
on a Ni-NTA affinity column followed by cleavage of the N-terminal affinity tag
C4LZN0, -
recombinant His-tagged enzyme from strain BL21 by nickel affinity chromatography
-
; recombinant His-tagged enzyme from Escherichia coli
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
on a glutathione-Sepharose resin
-
recombinant His6-tagged DRS-SUMO and DRS-ubiquitin fusion proteins by nickel affinity chromatography from Escherichia coli strain BL21(DE3), recombinant biotin-tagged DRS-ubiquitin fusion protein by avidin affinity chromatography from Escherichia coli strain Bl21(DE3)
-
wild-type enzyme and N-terminal 32-residue truncated form (hDRS delta 32), expressed in Escherichia coli as fusion proteins linked through a thrombin cleavage site with glutathione-S-transferase
-
recombinant His-tagged enzyme from Escherichia coli
-
on a glutathione-Sepharose resin
-, Q8I2B1
wild-type and two truncated forms
-
affinity chromatography
-
recombinant GST-tagged enzyme from Escherichia coli strain Top10 by glutathione affinity chromatography, the tag is then cleaved of by thrombin, and the enzyme is further purified by adsorption chromatography yielding an AspRS with a short additional amino acid stretch at its N-terminus
-
recombinant truncated enzyme from Escherichia coli
P04802
recombinant wild-type and mutants
-
wild-type and mutant enzymes
-
2 enzyme forms: PC-1, PC-2 and PC-3
-
enzyme form PC-1 (in addition 3 low MW enzyme forms exist: PC-2a, PC-2b and PC-2c)
-
; recombinant His-tagged enzyme from Escherichia coli
-
recombinant from overexpressing Escherichia coli strain JM103, to homogeneity
-
2 distinct enzyme forms: AspRS1 and AspRS2
-
recombinant AspRS2 from Escherichia coli by two different steps of anion exchange chromatography and hydroxy apatite chromatography
-
recombinant from overexpressing Escherichia coli
-
recombinant from overexpressing Escherichia coli, 34.3fold; recombinant from overexpressing Escherichia coli, 7.4fold
P36419, Q9LCY8
recombinant wild-type and seleno-Met isozyme 2 from overexpressing Escherichia coli
-
by Ni2+-nitrilotriacetic acid chromatography, cytosolic and mitochondrial fractions of Trypanosoma brucei are prepared
Q38CB4, Q585M3, -
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the full length sequence is cloned into the AVA0421 vector
C4LZN0, -
gene aspS, expression of the His-tagged enzyme in strain BL21
-
expression as His-tagged protein in Escherichia coli; expression in Escherichia coli
-
gene aspS, expression of His-tagged enzyme in Escherichia coli strain Bl21(DE3)
-
cloning into pQE70 vectors and expression in Escherichia coli TOP 10
-
expression in CHO cells
-
expression of His6-tagged or GST-tagged enzyme in Escherichia coli strain BL21(DE3), and expression of biotin- or His6-tagged DRS-SUMO and DRS-ubiquitin fusion proteins in Escherichia coli strain BL21(DE3), since the free DRS is expressed as an inactive and insoluble protein in Escherichia coli, SUMO is a small ubiquitin-like modifier molecule
-
gene DARS2, DNA and amino acid sequence determination and analysis
-
gene DARS2, located on chromosome 1, DNA and amino acid sequence determination and analysis, expression analysis, genotyping
-
into the vector pGEX-2T for expression in Escherichia coli ER2566 cells
-
wild-type enzyme and N-terminal 32-residues truncated form, expressed in Escherichia coli as fusion proteins linked through a thrombin cleavage site with glutathione-S-transferase
-
expression as His-tagged protein in Escherichia coli
-
into the vector pGEX-2T for expression in Escherichia coli ER2566 cells
-, Q8I2B1
overexpression in Escherichia coli
-
expression in Chinese hamster ovary cells
-
expression in Saccharomyces cerevisiae, wild-type enzyme and two truncated forms lacking 20 or 36 amino acid residues from their amino-terminal polypeptide extension
-
native recombinant AspRS from rat and the N-terminal truncated derivatives AspRS-DELTA20 and AspRS-DELTA36 expressed in yeast
-
expression in Saccharomyces cerevisiae, and expression as His-tagged protein
-
expression of wild-type and mutant enzymes in yeast YBC-603 cells, functional overexpression of GST- and His6-tagged enzyme in Escherichia coli strain Top10. Expression in yeast cells at low copy number, since overexpression is cytotoxic, while moderate AspRS accumulation in the cell is not deleterious
-
expression of wild-type enzyme, amino acid residues 1-557, and a truncated mutant enzyme, amino acid resdiues 71-557, as His-tagged proteins in Escherichia coli
-
mutants with substituted His residues expressed in Escherichia coli
-
overexpression of wild-type and mutants
-
truncated enzyme lacking 70 amino acid residues in the N-terminus, overexpression in Escherichia coli
P04802
expression as His-tagged protein in Escherichia coli; expression in Escherichia coli
-
expression in Escherichia coli
-
overexpression in Escherichia coli strain JM103
-
aspS1 gene, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strains DH5alpha and BL21; aspS2 gene, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strains DH5alpha and BL21
P36419, Q9LCY8
gene aspS2, expression in Escherichia coli strain JM103
-
overexpression in Escherichia coli
-
overexpression of wild-type isozyme AspRS2 and seleno-Met isozyme AspRS2 in Escherichia coli
-
the sequence corresponding to the carboxyl-terminal Ty1-tagged Tb-AspRS1 is cloned into a derivative of pLew-100 to allow tetracycline-inducible expression of the tagged protein in transgenic cell lines; the sequence corresponding to the carboxyl-terminal Ty1-tagged Tb-AspRS1 is cloned into a derivative of pLew-100 to allow tetracycline-inducible expression of the tagged protein in transgenic cell lines, into the vector pET15b for expression in Escherichia coli BL21-CodonPlus DE3-RIL cells
Q38CB4, Q585M3, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A471T
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143
D29N
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143
E118K
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143. 6.5fold increase in aminoacylation rate and 3fold decrease in amino acid activation reaction
E93K
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143
G90S
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143
G90V
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143
K198L
-
exchange of the conserved residue leads to a slightly increased binding of Asp, binding of Asn is strongly increased to the level of Asp, no ability to adenylate Asn, molecular dynamic simulation, binding mechansim and free energy
K198L
-
site-directed mutagenesis, the mutant shows reduced ATP-diphosphate exchange activity compared to the wild-type enzyme
K198L/D233E
-
slightly increased binding of Asp, binding of ASn is strongly increased to the level of Asp, no ability to adenylate Asn, molecular dynamic simulation, binding mechansim and free energy
K198L/Q199E/D233E
-
binding of Asp is strongly reduced, no ability to adenylate Asn, molecular dynamic simulation, binding mechansim and free energy
P555S
-
thermosensitive mutant, resulting in substitution of Pro 555 by Ser. Pro555Ser lowers the stability of the functional configuration of both the acylation and the amino acid activation sites but has no significant effect on substrate binding
Q199E
-
site-directed mutagenesis and molecular dynamics simulation of the mutation, reduction in binding free energy in the simulation in agreement with experiment, overview
Q199E
-
site-directed mutagenesis, the mutant shows reduced ATP-diphosphate exchange activity compared to the wild-type enzyme
R383C
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143
T89I
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143
C152F
-
the mutant shows 14fold reduced activity compared to the wild-type enzyme
D560V
-
the mutant shows 400fold reduced activity compared to the wild-type enzyme
hDRSDELTA32
-
N-terminal 32-residue truncated form, hDRSDELTA32, with lower thermal stability and ATP-diphosphate exchange activity, but higher aminoacylation activity. Fusion protein of glutathione-S-transferase and hDRSDELTA32 with lower thermal stability
Q184K
-
the mutant shows 170fold reduced activity compared to the wild-type enzyme
R263Q
-
the mutant shows 260fold reduced activity compared to the wild-type enzyme
D210A
-
central core binding mutant, increased acylation activity, increased dissociation constant
E188A
-
anticodon loop binding mutant, slightly reduced acylation activity, increased dissociation constant
E188A/S239A
-
anticodon-G73 binding mutant, reduced acylation activity, highly increased dissociation constant
E188A/T331A
-
anticodon-G73 binding mutant, highly reduced acylation activity, highly increased dissociation constant
E202A
-
central core binding mutant, slightly increased acylation activity, increased dissociation constant
E327A
-
acceptor arm binding mutant, reduced acylation activity
F127A
-
anticodon loop binding mutant, increased acylation activity, increased dissociation constant
F127A/D210A
-
anticodon binding mutant, reduced acylation activity, highly increased dissociation constant
F127A/E188A
-
anticodon binding mutant, reduced acylation activity, highly increased dissociation constant
F127A/S239A
-
anticodon-G73 binding mutant, reduced acylation activity, highly increased dissociation constant
F127A/T331A
-
anticodon-G73 binding mutant, highly reduced acylation activity, highly increased dissociation constant
F304A
-
terminal A binding mutant, reduced acylation activity
H116G
-
mutants with substituted His residues, His116Gly mutant with a slightly reduced rate of amino acid activation without affecting the other kinetic parameters, His271Gly mutant with completely destroyed activity, His332Gly mutant with 60% decrease in rate of tRNA aminoacylation and no significant changes in the other parameters, His334Gly mutant with 70% decrease in amino acid activation, complete loss of tRNA aspartylation and slightly increased Km for ATP, His271Ala mutant with 25% decrease in the rate of tRNA charging. His334 seems do be part of the active site, while His271 and His332 play an important structural role
H334A
-
acceptor arm binding mutant, highly reduced acylation activity, increased dissociation constant
K142A
-
anticodon loop binding mutant, increased acylation activity, increased dissociation constant
K142A/E188A
-
anticodon binding mutant, reduced acylation activity, highly increased dissociation constant
K155A
-
central core binding mutant, increased acylation activity, increased dissociation constant
K180A
-
anticodon loop binding mutant, slightly increased acylation activity, increased dissociation constant
K293A
-
acceptor arm binding mutant, highly increased acylation activity, increased dissociation constant
K428A
-
acceptor arm binding mutant, increased acylation activity, increased dissociation constant
N117A
-
anticodon loop binding mutant, increased acylation activity, increased dissociation constant
N227A
-
central core binding mutant, slightly increased acylation activity, increased dissociation constant
N328A
-
acceptor arm binding mutant, reduced acylation activity, increased dissociation constant
N328A/S329A/T331A
-
G73 binding mutant, highly reduced acylation activity, increased dissociation constant
P273G
-
Pro273Gly mutant. Catalytic properties of native and Pro273Gly homodimers or heterodimers of AspRS molecules, confirm the participation of Pro273 in subunit association
Q121A
-
anticodon loop binding mutant, increased acylation activity, increased dissociation constant
Q138A
-
anticodon loop binding mutant, slightly increased acylation activity, increased dissociation constant
Q138A/E188A
-
anticodon binding mutant, reduced acylation activity, highly increased dissociation constant
Q138A/R119A
-
anticodon binding mutant, reduced acylation activity, highly increased dissociation constant
Q300A
-
terminal A binding mutant, reduced acylation activity
R119A
-
anticodon loop binding mutant, increased acylation activity, increased dissociation constant
R119A/E188A
-
anticodon binding mutant, highly reduced acylation activity, highly increased dissociation constant
R485K
-
site-directed mutagenesis, the substitution in the catalytic site completely inhibits aspartylation by impairing ATP binding, this mutant still retains the capacity to be modified and shows the same pattern as wild-type AspRS on the two-dimensional gel, thus the modifications are not the result of autoaspartylation
S181A
-
anticodon loop binding mutant, increased acylation activity, increased dissociation constant
S280A
-
terminal A binding mutant, reduced acylation activity
S301A
-
terminal A binding mutant, reduced acylation activity, slightly increased dissociation constant
S329A
-
acceptor arm binding mutant, increased acylation activity, increased dissociation constant
S423A
-
acceptor arm binding mutant, reduced acylation activity
T124A
-
anticodon loop binding mutant, slightly reduced acylation activity, increased dissociation constant
T230A
-
central core binding mutant, increased acylation activity, increased dissociation constant
T331A
-
acceptor arm binding mutant, reduced acylation activity, increased dissociation constant
T424A
-
acceptor arm binding mutant, acylation activity similar to the wild-type, increased dissociation constant
W26H
-
discriminating AspRS gains the ability to form Asp-tRNAAsn. Mutation causes a 1.5fold decrease in overall catalytic efficiency for Asp-tRNAASp synthesis; the wild-type enzyme shows no activity with tRNAAsn, the mutant is active with tRNAAsn
L30F
-
mutant enzyme is still able to aminoacylate the native tRNAAsp in vivo at a level sufficient to complement the defective strain CS143
additional information
-
transplantation of the L45 loop of Escherichia coli AspRS and two residues flanking the loop inside the Escherichia coli lysyl-tRNALys ligase sequence causes a loss of lysyl-tRNALys ligase capacity to aminoacylate tRNALys. The chimeric enzyme acquires the capacity to charge tRNAAsp with lysine
L626V
-
the mutant shows 210fold reduced activity compared to the wild-type enzyme
additional information
-
enzyme deficiency leads to autosomal recessive disease leukoencephalopathy with brain stem and spinal cord involvement and lactate elevation, i.e. LBSL, often manifesting in early childhood, affected individuals develop slowly progressive cerebellar ataxia, spasticity and dorsal column dysfunction, sometimes with a mild cognitive deficit or decline, phenotype, mutational analysis, overview
additional information
-
a male patient with two mutations heterogenous in the DARS2 gene, shows cerebellar, pyramidal and dorsal column dysfunctions and specific magnetic resonance imaging and characteristic magnetic resonance spectroscopy abnormalities, leukoencephalopathy with brain stem and spinal cord involvement, phenotype, overview; two deletion mutations in the DARS2 gene cause the autosomal recessive disorder leukoencephalopathy with brainstem and spinal cord involvement and lactate elevation. determination of cerebellar, pyramidal and dorsal column dysfunctions and abnormalities, e.g. in in the superior cerebellar peduncles, the intraparenchymal trajectories of the trigeminal nerves, the pyramidal tracts, and the medial lemniscus, by resonance imaging, MRI, and magnetic resonance spectroscopy, MRS, phenotype, overview
S45G
Q6PI48
a naturally occuring mutation identified in patients suffering leukoencephalopathy with brain stem and spinal cord involvement and lactate elevation. The mutant enzyme is not processed due to nontranslocation of the protein
additional information
-, Q8I2B1
the RNA binding motif is mutated, from 70KKKEKKAKK78 to 70QEEREEGQM78
Asp-DELTA20
-
native recombinant AspRS from rat and the N-terminal truncated derivatives Asp-DELTA20 and AspRS-DELTA36 expressed in yeast. A moderate but significant drop in affinity towards the multisynthetase complex is inferred by the removal of the N-terminal domain. This domain is absolutely required in vivo for association within the multisynthetase structure
additional information
-
mutant enzyme with a deletion of 34 amino acids from its N-terminus does not associate within the complex from Chinese hamster ovary cells. A chimeric enzyme made of the amino-terminal moiety of rat liver aspartyl-tRNA synthetase fused to the catalytic domain of yeast lysyl-tRNA synthetase, expressed in Lys-101 cells (a Chinese hamster ovary cell line with a temperature-sensitive lysyl-tRNA synthetase) does not associate within the multisynthetase complex and cannot restore normal growth of mutant cells
additional information
-
two truncated forms lacking 20 or 36 amino acid residues from their amino-terminal polypeptide extension, expressed in Saccharomyces cerevisiae, are less stable than wild-type enzyme
K553A
-
acceptor arm binding mutant, acylation activity similar to wild-type, increased dissociation constant
additional information
-
C-terminal and N-terminal truncated forms. On the C-terminal side, very limited modifications readily affect the enzyme properties. The N-terminal sequence up to amino acid 70 is dispensable for activity, domains beyond amino acid 70 have increasing catalytic importance
additional information
-
truncated form that has lost the first 50-64 residues, with full retention of both the activity and the dimeric structure
additional information
-
cooperativity analysis of the G73 and anticodon binding interactions, overview, effect of mutations on cell growth
additional information
-
construction of several N-terminal deletion mutants with altered mRNA binding properties, overview
additional information
-
proteomic analysis of mutant cells in comparison to wild-type cells
K85P
-
discriminating AspRS gains the ability to form Asp-tRNAAsn. Mutation impairs the ability to synthesize Asp-trNAASp in vitr, 8fold increase in KM-value for tRNAAsp; the wild-type enzyme shows no activity with tRNAAsn, the mutant is active with tRNAAsn
additional information
-
wild-type discriminating enzyme is engineered to a non-discriminating mutant by site-directed mutagenesis, a L1 loop exchange
W26H/K85P
-
discriminating AspRS gains the ability to form Asp-tRNAAsn; the wild-type enzyme shows no activity with tRNAAsn, the mutant is active with tRNAAsn
additional information
-
construction of 7 mutants for crystallization
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
denaturation by 4 M urea, 30% recovery after dialysis
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
the adaption of pathogens to antibiotics calls for new target macromolecules and new types of inhibitors
pharmacology
-
the enzyme is the target of peptide nucleotide antibiotic Microcin C
medicine
-
the adaption of pathogens to antibiotics calls for new target macromolecules and new types of inhibitors
medicine
P14868
for designing selective inhibitors of yeast AspAS, well distinguished from mammalian AspRS, the understanding of the structural features is necessary
medicine
-, Q3U987
for designing selective inhibitors of yeast AspAS, well distinguished from mammalian AspRS, the understanding of the structural features is necessary
medicine
-
the adaption of pathogens to antibiotics calls for new target macromolecules and new types of inhibitors