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6.1.1.4: leucine-tRNA ligase

This is an abbreviated version!
For detailed information about leucine-tRNA ligase, go to the full flat file.

Word Map on EC 6.1.1.4

Reaction

ATP
+
L-leucine
 +
tRNALeu
=
AMP
 +
diphosphate
 +
L-leucyl-tRNALeu

Synonyms

AaLeuRS, alphabeta-LeuRS, b0642, cytoplasmic LeuRS, EcLeuRS, GlLeuRS, HcleuRS, hs mt LeuRS, JW0637, LARS, LARS1, LARS2, Leucine translase, Leucine--tRNA ligase, Leucyl-transfer ribonucleate synthetase, Leucyl-transfer ribonucleic acid synthetase, Leucyl-transfer RNA synthetase, leucyl-tRNA ligase, leucyl-tRNA syntethase, Leucyl-tRNA synthetase, leucyl-tRNA synthetase 1, leucyl—tRNA synthetase, LeuRS, LeuRS1, LeuRS2, LeuRSTT, leuS, LRS, MmLeuRS, More, mt leucyl-tRNA synthetase, mt-LeuRS, mtLeuRS, PhLeuRS, Synthetase, leucyl-transfer ribonucleate, ycLeuRS

ECTree

     6 Ligases
         6.1 Forming carbon-oxygen bonds
             6.1.1 Ligases forming aminoacyl-tRNA and related compounds
                6.1.1.4 leucine-tRNA ligase

Engineering

Engineering on EC 6.1.1.4 - leucine-tRNA ligase

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A156V
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is identical to the wild-type ratio
D373A
-
mutant defective in post-transfer editing function
G237D
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 1.7fold higher than the wild-type ratio
K238A
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 2.4fold higher than the wild-type ratio
K599A
-
site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K599A/K600A
-
site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K599A/K600A/K605A/K606A
-
site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K600A
-
site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K605A
-
site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K605A/K606A
-
site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K606A
-
site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
L283F
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 2fold higher than the wild-type ratio
M159A
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 80% of the wild-type value
M160N
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 2.4fold higher than the wild-type ratio
N152A
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 16% of the wild-type ratio
N163A
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 2.4fold higher than the wild-type ratio
Q234H
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is identical to the wild-type ratio
Q260stop
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 0.02% of the wild-type ratio
R94A/R98A
site-directed mutagenesis, 83% reduced catalytic efficiency compared to wild-type
R94E
site-directed mutagenesis, mutating Arg94 to Glu decreases kcat/Km values to 22% of that of wild-type AaLeuRS
R94E/R98E
site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type
R98A
site-directed mutagenesis, the mutation does not alter the catalytic efficiency
R98E
site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type
T273R
-
no change in aminoacylation activity, but the deacylation of Ile-tRNALeu is strongly impaired. Mutant still exhibits 70% of wild-type AMP formation
V286stop
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 0.6% of the wild-type ratio
ts025C1
-
temperature sensitive mutant ts025C1
tsH1
-
temperature sensitive mutant tsH1
A293D
A293E
-
site-directed mutagenesis, the mutant activity is similar to the wild-type enzyme
A293F
-
54% decreased activity compared to the wild-type, more sensitive too inhibition by ATP
A293G
-
50% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP
A293I
-
51% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP
A293K
-
site-directed mutagenesis, the post-transfer editing activity of the isolated CP1-domain is enhanced compared to the wild-type enzyme's domain
A293R
A293Y
-
50% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP
C159A
site-directed mutagenesis, structure comparison with the wild-type
C176A
site-directed mutagenesis, structure comparison with the wild-type
C179A
site-directed mutagenesis, structure comparison with the wild-type
D251W
-
site-directed mutagenesis, editing site mutant, the substrate specificity and charging fidelity is retained
D342A
site-directed mutagenesis, the mutant shows altered deacylation activity with amino acids norvaline, isoleucine, and leucine compared to the wild-type enzyme, overview
D345A
DELTA788-798
-
partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. This mutant shows almost no aminoacylation activity. Mutant shows reduced deacylation activity against mischarged Ile-tRNALeu
DELTA790-798
-
partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant shows reduced deacylation activity against mischarged Ile-tRNALeu
DELTA792-798
-
partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu
DELTA793
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA794
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA794-798
-
partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu
DELTA795
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA795-796
-
two-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA795-798
-
partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu
DELTA796
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA796-798
DELTA797
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA797-798
-
two-site deletion at the more flexible end of the peptide linker: mutant exhibits lower aminoacylation activity compared to wild-type
E184A
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
E184R
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme. The substitution specifically inhibits tRNA-dependent pre-transfer editing
E184R/T252R
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
E292A
-
unaltered specific activity in amino acid activation reaction, 61% reduced aminoacylation activity compared to the wild-type
E292D
-
unaltered specific activity in amino acid activation reaction, 53% reduced aminoacylation activity compared to the wild-type
E292F
-
unaltered specific activity in amino acid activation reaction, 60% reduced aminoacylation activity compared to the wild-type
E292K
-
unaltered specific activity in amino acid activation reaction, 85% reduced aminoacylation activity compared to the wild-type
E292Q
-
unaltered specific activity in amino acid activation reaction, 54% reduced aminoacylation activity compared to the wild-type
E292S
-
unaltered specific activity in amino acid activation reaction, 66% reduced aminoacylation activity compared to the wild-type
E797GGG
-
mutant shows no altered aminoacylation activity compared to wild-type
E797PPP
-
mutant shows no altered aminoacylation activity compared to wild-type
G225P
-
abolishes tRNA leucylation due to a defect in leucine activation, decrease in deacylation of Ile-tRNALeu
G229P
-
increased aminoacylation activity compared to the wild-type, mutant deacylates Ile-tRNALeu similar to wild-type
G229P/T252A
-
double mutant rescues leucylation activity to levels comparable to wild-type and retains deacylation activity of LeutRNALeu that is characteristic of the T252A mutation
G407P
-
aminoacylates tRNALeu and decylates Ile-tRNALeu as well as wild-type
G409P
-
increased aminoacylation activity compared to the wild-type, mutant deacylates Ile-tRNALeu similar to wild-type
G409P/T252A
-
double mutant fails to rescue the T252A mutation in LeuRS
K671A
site-directed mutagenesis, the mutation does no affect the catalytic efificiency
K809A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
K846A
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
K846A/K853A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
K846E
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
K846E/K853E
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
K853A
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-typ enzyme
K853E
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
L570F
-
the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, it shows a catalytic turnover for isoleucine decreased by a factor of 2, L570F has an 11fold higher Km for leucine compared to the wild-type enzyme, the activity is reduceDdcompared to the wild-type enzyme
L570K
-
the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, it shows a catalytic turnover for isoleucine decreased by a factor of 2, L570F has an 11fold higher Km for leucine compared to the wild-type enzyme, the activity is reduced compared to the wild-type enzyme
L570R
-
the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, it shows a catalytic turnover for isoleucine decreased by a factor of 2, L570R has a 4fold stronger binding affinity for leucine compared to the wild-type enzyme, the activity is reduce compared to the wild-type enzyme
L854A
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
L855A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
M328K
-
7% increased activity compared to the wild-type
M336A
-
site-directed mutagenesis, editing site mutant, the mutant shows a small increase in leucine editing activity
M336F/T252A
-
site-directed mutagenesis, editing site mutant, the T252A mutation uncouples specificity, M336F/T252A double LeuRS mutant exhibited only slightly increased leucylation activity relative to the T252A single mutation
N807A
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
N807A/N856A
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
N856A
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
Q805A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
Q805A/N807A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
Q805A/N807A/N856A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
R185E
the mutation significantly enhances tRNA-dependent pre-transfer editing activity
R249F
-
site-directed mutagenesis, editing site mutant, editing activity of Leu-tRNALeu is decreased
R249F/T252A
-
site-directed mutagenesis, editing site mutant, the T252A mutation uncouples specificity
R249T
-
site-directed mutagenesis, editing site mutant, the mutant shows increased activity with tRNALeu, but even higher activity with tRNAIle compared to the wild-type enzyme
R249T/D251W
-
site-directed mutagenesis, editing site mutant, the mutant shows decreased hydrolysis of mischarged Ile-tRNALeu compared to the wild type enzyme
R286E
the mutation significantly enhances tRNA-dependent pre-transfer editing activity
R344A
site-directed mutagenesis, the mutant shows altered deacylation activity with amino acids norvaline, isoleucine, and leucine compared to the wild-type enzyme, overview
R668A
site-directed mutagenesis, the mutant shows 77% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type
R668A/R672A
site-directed mutagenesis, the mutant shows 93.6% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type
R668E
site-directed mutagenesis, the mutant shows 95% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type
R668E/R672E
site-directed mutagenesis, the mutant shows 98.6% reduced catalytic efficiency compared to wild-type. But the almost inactive mutant exhibits intact Leu activation activity comparable with the wild-type enzyme
R672A
site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type
R672E
site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type
R811A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
T247A/T248A
-
533fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247S/T248S
-
77fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247V
T247V/T248V
T248A
site-directed mutagenesis, the mutant shows altered deacylation activity with amino acids norvaline, isoleucine, and leucine compared to the wild-type enzyme, overview
T248V
T252A
T252D
-
mutation results in isoleucylation of tRNALeu, editing activity is impaired, ATP hydrolysis in presence of norvaline is 27% of the wild-type value, ATP hydrolysis in presence of leucine is 98% of the wild-type value
T252E
T252E/M328K
-
activity is similar to the wild-type
T252F
-
impaired proofreading mechanism, increase rate of misaminoacylation with isoleucine and valine
T252G
-
the mutant enzyme, like the native enzyme, does not mischarge tRNALeu with isoleucine, ATP hydrolysis in presence of norvaline is 2.1fold higher than wild-type value, ATP hydrolysis in presence of leucine is 60% of the wild-type value
T252L
-
impaired proofreading mechanism, increase rate of misaminoacylation with isoleucine and valine
T252R
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
T252S
-
the ratio of turnover-number to KM-value for L-leucine in aminoacylation is 60% of the wild-type ratio, the ratio of turnover-number to KM-value for tRNALeu in aminoacylation is 80% of the wild-type ratio. The ratio of turnover-number to Km value for Leu-tRNALeu(UAA) is 7.5fold higher than the wild-type value, the ratio of turnover-number to Km value for Ile-tRNALeu(UAA) is 1.6fold lower than the wild-type value
T252V
-
the ratio of turnover-number to KM-value for L-leucine in aminoacylation is identical to wild-type ratio, the ratio of turnover-number to KM-value for tRNALeu in aminoacylation is 90% of the wild-type ratio. The ratio of turnover-number to Km value for Leu-tRNALeu(UAA) is 1.75fold higher than the wild-type value, the ratio of turnover-number to Km value for Ile-tRNALeu(UAA) is 13.3fold higher than the wild-type value
T252Y
T272R
-
no change in aminoacylation activity, but the deacylation of Ile-tRNALeu is strongly impaired. Mutant still exhibits 45% of wild-type AMP formation
V338A
-
site-directed mutagenesis, editing site mutant, it shows increased post-transfer editing activity of Leu-tRNALeu compared to the wild-type enzyme
V338D
-
site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
V338E
-
site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
V338F
-
site-directed mutagenesis, editing site mutant, single introduction of the bulky phenylalanine residue nearly abolished post-transfer editing activity and facilitated mischarging of both isoleucine and valine to tRNALeu, 3000fold reduced activity
V338F/T252A
-
site-directed mutagenesis, editing site mutant, the T252A mutation uncouples specificity
V338L
-
site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
D173A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
D444A
D444E
-
site-directed mutagenesis
D588A
-
kcat/Km: 0.57 (ATP), 0.37 (Leu), mutant displays lower amino acid activation and aminoacylation activities than wild-type
D603A
-
kcat/Km: 0.91 (ATP), 0.94 (Leu), similar activity compared to wild-type
DELTA581-617
-
deletion of the CP2 domain, an insertion domain called connective peptide 2, of Giardia lamblia LeuRS shows that the CP2 domain is indispensable for amino acid activation, post-transfer editing and contributes to LeuRS-tRNALeu binding affinity. CP2 domain of Pyrococcus horikoshii LeuRS but not that of Escherichia coli LeuRS can partially restore amino acid activation and post-transfer editing functions suggesting that the functions of the CP2 domain are dependent on its location in the primary sequence of LeuRS
E165A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
E167A
-
site-directed mutagenesis, the mutant shows defects in leucine activation, mutant kinetics compared to the wild-type enzyme, overview
E298A
-
activity similar to wild-type
F171A
-
site-directed mutagenesis, inactive mutant
K139A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K141A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K142A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K144A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K148A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K152A
-
site-directed mutagenesis, the mutant shows defects in leucine activation, mutant kinetics compared to the wild-type enzyme, overview
K166A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K170A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K299A
-
activity similar to wild-type
K303A
-
activity similar to wild-type
K587A
-
kcat/Km: 0.43 (ATP), 0.42 (Leu), mutant displays lower amino acid activation and aminoacylation activities than wild-type
K606A
-
mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities
K606D
-
kcat/Km: 0.86 (ATP), 0.83 (Leu), similar leucine activation and post-transfer editing activities compared to wild-type
K606E
-
kcat/Km: 0.90 (ATP), 0.99 (Leu), no difference in leucine activation and post-transfer editing activities compared to wild-type
K606L
-
kcat/Km: 0.85 (ATP), 0.98 (Leu), similar leucine activation and post-transfer editing activities compared to wild-type
K606R
-
kcat/Km: 0.98 (ATP), 0.99 (Leu), no difference in leucine activation and post-transfer editing activities compared to wild-type
N301A
-
activity similar to wild-type
Q154A
-
site-directed mutagenesis, the mutant shows defects in leucine activation, mutant kinetics compared to the wild-type enzyme, overview
R338A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S153A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
S295A
-
activity similar to wild-type
T341A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T341R
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
W155A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
W586A
-
mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities
Y515A
-
109% of wild-type activity
Y515E
-
42% of wild-type activity
Y515K
-
51% of wild-type activity
Y520A
-
68% of wild-type activity
Y520E
-
41% of wild-type activity
Y520H
-
46% of wild-type activity
Y581A
-
mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities
Y581E
-
mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity
Y581F
-
mutant shows some but significantly reduced leucine activation activity its post-transfer editing activity is similar to wild-type
Y581K
-
mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity
Y581S
-
mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity
A3243G
-
respiratory chain defects in A3243G mutant cells is suppressed by overexpressing human mitochondrial leucyl-tRNA synthetase. The rates of oxygen consumption in suppressed cells are directly proportional to the levels of leucyl-tRNA synthetase. 15fold higher levels of leucyl-tRNA synthetase results in wild-type respiratory chain function. The suppressed cells have increased steady-state levels of tRNA(Leu(UUR)) and up to 3fold higher steady-state levels of mitochondrial translation products, but do not have rates of protein synthesis above those in parental mutant cells
A525S
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
C527E
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
D250A
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
D250E
site-directed mutagenesis, the mutant shows slightly altered kinetics and slightly reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
D250N
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
D250R
site-directed mutagenesis, inactive mutant
D252R
site-directed mutagenesis, inactive mutant
D399A
D399K
-
mutant is resitant to inhibitor 5-fluoro-2,1-benzoxaborol-1(3H)-ol but more sensitive to norvaline inhibition
D528R
site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 85% reduced amino acid activation activity compared to the wild-type enzyme
F50A/Y52A
site-directed mutagenesis, the leucine-binding deficient LRS mutant also activates Vps34, but to a lesser degree and in a leucine-independent manner
G245A
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
G245D
site-directed mutagenesis, the mutant shows altered kinetics and 50% reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
G245P
site-directed mutagenesis, inactive mutant
G245R
site-directed mutagenesis, the mutant shows altered kinetics and 50% reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
H251D
site-directed mutagenesis, inactive mutant
K600F
-
the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, the mutant demonstrates a 9fold decrease in its ability to distinguish between leucine and isoleucine effectively, the activity is reduced compared to the wild-type enzyme
K600L
-
the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, the mutant demonstrates an 11fold increase in its ability to distinguish between leucine and isoleucine effectively, the activity is reduced compared to the wild-type enzyme
K600R
-
the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, the mutant shows a slight decrease in activity compared to the wild-type enzyme
P242E
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
P247A
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
Q529A
site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 70% reduced amino acid activation activity compared to the wild-type enzyme
R236D
site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 30% reduced amino acid activation activity compared to the wild-type enzyme
R517D
site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 90% reduced amino acid activation activity compared to the wild-type enzyme
R766A
site-directed mutagenesis, the mutation decreases the kcat/Km value to less than 10% that of the wild-type enzyme hcLeuRS
S519G
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
T298A
activity similar to wild-type, mutation maintains Ile-tRNALeu deacylation activity
T298Y
mutation uncouples specificity in the editing active site and mutant hydrolyzes Leu-tRNALeu
V523I
site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme
W530A
site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 50% reduced amino acid activation activity compared to the wild-type enzyme
Y240A
site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 50% reduced amino acid activation activity compared to the wild-type enzyme
Y531A
site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 50% reduced amino acid activation activity compared to the wild-type enzyme
Y534A
site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 60% reduced amino acid activation activity compared to the wild-type enzyme
K452A
Mesomycoplasma mobile
site-directed mutagenesis, the mutation has only a minimal effect on aminoacylation activity, the Km values is not significantly altered compared to wild-type
K452E
Mesomycoplasma mobile
site-directed mutagenesis, the mutation has only a minimal effect on aminoacylation activity, the Km values is not significantly altered compared to wild-type
K598A
Mesomycoplasma mobile
-
the mutation simultaneously reduces the tRNA-binding strength and aminoacylation and editing capacities of the enzyme's leucine-specific domain
R456A
Mesomycoplasma mobile
site-directed mutagenesis, 75% reduced catalytic efficiency compared to wild-type, the Km values is not significantly altered
R456E
Mesomycoplasma mobile
site-directed mutagenesis, 79% reduced catalytic efficiency compared to wild-type, the Km values is not significantly altered
K452A
Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711
-
site-directed mutagenesis, the mutation has only a minimal effect on aminoacylation activity, the Km values is not significantly altered compared to wild-type
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K452E
Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711
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site-directed mutagenesis, the mutation has only a minimal effect on aminoacylation activity, the Km values is not significantly altered compared to wild-type
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R456A
Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711
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site-directed mutagenesis, 75% reduced catalytic efficiency compared to wild-type, the Km values is not significantly altered
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R456E
Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711
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site-directed mutagenesis, 79% reduced catalytic efficiency compared to wild-type, the Km values is not significantly altered
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L949A
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the mutant shows 9.5% activity compared to the wild type enzyme
L949K
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the mutant shows 2.6% activity compared to the wild type enzyme
L964A
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the mutant shows 85% activity compared to the wild type enzyme
L964K
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the mutant shows 8.6% activity compared to the wild type enzyme
Q915A
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the mutant shows 99% activity compared to the wild type enzyme
Q915K
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the mutant shows 54% activity compared to the wild type enzyme
R921A
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the mutant shows 37% activity compared to the wild type enzyme
R921K
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the mutant shows 83% activity compared to the wild type enzyme
V910A
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the mutant shows 90% activity compared to the wild type enzyme
V910P
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the mutant shows 3.7% activity compared to the wild type enzyme
V910W
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the mutant shows 93% activity compared to the wild type enzyme
Q915A
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the mutant shows 99% activity compared to the wild type enzyme
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Q915K
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the mutant shows 54% activity compared to the wild type enzyme
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V910A
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the mutant shows 90% activity compared to the wild type enzyme
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V910P
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the mutant shows 3.7% activity compared to the wild type enzyme
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V910W
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the mutant shows 93% activity compared to the wild type enzyme
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D121A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
D332A
D98A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
E113A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
E114A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
F119A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
I104A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
I115A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
K100A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
K100A/Y105A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
K100A/Y109A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
K692A
site-directed mutagenesis, the mutation has no effect on tRNA charging activity
K696A
site-directed mutagenesis, the mutant shows a highly reduced kcat value compared to wild-type, while the Km value is 3fold increased
K699A
site-directed mutagenesis, the mutation has no effect on tRNA charging activity
N96A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
R106A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
R698A
site-directed mutagenesis, the mutation has no effect on tRNA charging activity
R703A
site-directed mutagenesis, kcat of mutant PhLeuRSR703A is much lower than that of wild-type PhLeuRS
R97A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
T101A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
T118A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
V108A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
W103A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
Y105A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
Y109A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
K692A
Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3
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site-directed mutagenesis, the mutation has no effect on tRNA charging activity
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K696A
Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3
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site-directed mutagenesis, the mutant shows a highly reduced kcat value compared to wild-type, while the Km value is 3fold increased
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K699A
Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3
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site-directed mutagenesis, the mutation has no effect on tRNA charging activity
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R698A
Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3
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site-directed mutagenesis, the mutation has no effect on tRNA charging activity
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R703A
Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3
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site-directed mutagenesis, kcat of mutant PhLeuRSR703A is much lower than that of wild-type PhLeuRS
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D357A
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site-directed mutagenesis, the mutant shows reduced activity and abolished editing activity and misaminoacylated isoleucine to tRNALeu compared to the wild-type enzyme
D418R
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
D419A
DELTA270-530
deletion of the CP1 domain shows that the mutant is not able to rescue LeuRS knock-out strain
DELTA314-319
deletion of the T-rich region shows that the mutant is able to rescue LeuRS knock-out strain with a grwoth rate similar to wild-type
DELTA819-828
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deletion of the C-terminal domain peptide linker stimulates aminoacylation and editing activity shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu
K404Y
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
R265A
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site-directed mutagenesis, the mutant shows reduced activity and abolished post-transfer editing activity compared to the wild-type enzyme
R449A
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site-directed mutagenesis, nearly inactive mutant
R449E
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
R449K
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320, 30fold reduced activity compared to the wild-type enzyme
R451A
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site-directed mutagenesis, nearly inactive mutant
R451E
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
R451K
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site-directed mutagenesis, mutation within the RDW peptide, complementation of the null mutant strain QBY320, 11fold reduced activity compared to the wild-type enzyme
S416D
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
T263V/T264V
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site-directed mutagenesis, the mutant shows reduced activity and decreased post-transfer editing activity compared to the wild-type enzyme
T31E
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Mutant T319A shows mischarging capacity with Ile. Growth rate is severly inhibited in medium containing a large excess of norvaline and reduced leucine
T347A
mutant is inhibited by AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
T410A
mutant is inhibited by AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
W445A
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site-directed mutagenesis, nearly inactive mutant
W445F
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445H
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445K
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445Y
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site-directed mutagenesis, mutation within the RDW peptide, weak complementation of the null mutant strain QBY320, 30fold reduced activity compared to the wild-type enzyme
D347A
mutation of the highly conserved Asp residue, located in the CP1 domain, is responsible for editing mechanism, slightly reduced activity with L-leucine, mutant mischarges tRNALeu with isoleucine
additional information