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Information on EC 6.1.1.6 - lysine-tRNA ligase and Organism(s) Escherichia coli and UniProt Accession P0A8N3
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6.1.1.6 lysine-tRNA ligaseSpecify your search results
Word Map
- 6.1.1.6
- synthetases
- aminoacylation
- lysylation
-
anticodon
-
aarss
-
isoacceptors
-
diadenosine
- aspartyl-trna
-
gagpol
- tetraphosphate
- trnalys3
- prors
- valrs
- leurs
- arginyl-trna
-
isoleucyl-trna
- glnrs
-
atp-ppi
-
anticodon-binding
-
prolyl-trna
-
multi-trna
- asprs
- diagnostics
- pharmacology
- medicine
- drug development
- synthesis
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
lysyl-trna synthetase, lysrs, lysrs1, lysrs2, lysyl trna synthetase, mitochondrial lysyl-trna synthetase, cytoplasmic lysyl-trna synthetase, class i lysrs, premsk1p, pfkrs, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
L-Lysine-transfer RNA ligase
Lysine translase
Lysine--tRNA ligase
Lysine-tRNA synthetase
LysRS
LysU
Lysyl-transfer ribonucleate synthetase
Lysyl-transfer RNA synthetase
Lysyl-tRNA synthetase
Synthetase, lysyl-transfer ribonucleate
L-Lysine-transfer RNA ligase
-
-
-
-
Lysine translase
-
-
-
-
Lysine--tRNA ligase
-
-
-
-
Lysine-tRNA synthetase
-
-
-
-
LysRS
-
-
-
-
Lysyl-transfer ribonucleate synthetase
-
-
-
-
Lysyl-transfer RNA synthetase
-
-
-
-
Lysyl-tRNA synthetase
-
-
-
-
Lysyl-tRNA synthetase
-
-
Synthetase, lysyl-transfer ribonucleate
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-lysine + tRNALys = AMP + diphosphate + L-lysyl-tRNALys
active site structure and substrate binding
ATP + L-lysine + tRNALys = AMP + diphosphate + L-lysyl-tRNALys
active site structure
-
ATP + L-lysine + tRNALys = AMP + diphosphate + L-lysyl-tRNALys
two-step reaction mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
esterification
Aminoacylation
esterification
-
-
-
-
Aminoacylation
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
L-lysine:tRNALys ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY
9031-26-9
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + L-lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
-
?
AMP + diphosphate + L-methionyl-tRNALys
ATP + L-methionine + tRNALys
-
-
-
-
r
AMP + L-threonyl-tRNALys + diphosphate
ATP + L-threonine + tRNALys
-
-
-
-
r
ATP + 4-aminobutanoate + tRNALys
AMP + 4-aminobutyryl-tRNALys + diphosphate
-
-
-
?
ATP + L-alanine + tRNALys
AMP + L-alanyl-tRNALys + diphosphate
-
264000fold lower activity than with L-lysine, deacylation of the mischarged tRNALys, addition of 2 mM L-lysine abolishes the reaction
-
r
ATP + L-cysteine + tRNALys
AMP + L-cysteinyl-tRNALys + diphosphate
-
750000fold lower activity than with L-lysine, deacylation of the mischarged tRNALys, addition of 2 mM L-lysine abolishes the reaction
-
r
ATP + L-glutamate + tRNALys
AMP + L-glutamyl-tRNALys + diphosphate
-
low activity
-
?
ATP + L-leucine + tRNALys
AMP + L-leucyl-tRNALys + diphosphate
-
132000fold lower activity than with L-lysine, addition of 2 mM L-lysine abolishes the reaction
-
r
ATP + L-lysinamide + tRNALys
AMP + L-aminolysyl-tRNALys + diphosphate
-
low activity
-
?
ATP + L-lysine
?
-
a small fraction of lysine is converted to lysine lactam in absence or presence of tRNALys
-
?
ATP + L-lysine + Borellia burgdorferi tRNALys
AMP + diphosphate + L-lysyl-tRNALys
-
-
-
-
?
ATP + L-lysine + Escherichia coli G2.U71 tRNALys
AMP + diphosphate + L-lysyl-tRNALys
-
-
-
-
r
ATP + L-lysine + Escherichia coli wild type tRNALys
AMP + diphosphate + L-lysyl-tRNALys
-
-
-
-
r
ATP + L-lysine + tRNALysCUU
AMP + L-lysyl-tRNALysCUU + diphosphate
-
-
-
-
?
ATP + L-lysine + tRNALysUUU
AMP + L-lysyl-tRNALysUUU + diphosphate
-
-
-
-
?
ATP + L-lysine + tRNATyrCUA
AMP + L-lysyl-tRNATyrCUA + diphosphate
-
has a weak activity to tRNATyrCUA with L-lysine
-
-
?
ATP + L-lysine ethyl ester + tRNALys
AMP + ethyl-L-lysyl-tRNALys + diphosphate
-
-
-
?
ATP + L-lysine hydroxamate + tRNALys
AMP + L-lysine hydroxamoyl-tRNALys + diphosphate
-
-
-
?
ATP + L-lysine methyl ester + tRNALys
AMP + methyl-L-lysyl-tRNALys + diphosphate
-
-
-
?
ATP + L-methionine + tRNALys
AMP + L-methionyl-tRNALys + diphosphate
-
32000fold lower activity than with L-lysine, addition of 2 mM L-lysine abolishes the reaction
-
r
ATP + L-ornithine
?
-
L-ornithine is converted to ornithine lactam in absence or presence of tRNALys
-
?
ATP + L-serine + tRNALys
AMP + L-seryl-tRNALys + diphosphate
-
562000fold lower activity than with L-lysine, deacylation of the mischarged tRNALys, addition of 2 mM L-lysine abolishes the reaction
-
r
ATP + L-threonine + tRNALys
AMP + L-threonyl-tRNALys + diphosphate
-
16000fold lower activity than with L-lysine, addition of 2 mM L-lysine abolishes the reaction
-
r
ATP + ornithine + tRNALys
AMP + ornithyl-tRNALys + diphosphate
-
-
-
?
ATP + L-arginine + tRNALys
AMP + L-arginyl-tRNALys + diphosphate
-
low activity
-
?
ATP + L-arginine + tRNALys
AMP + L-arginyl-tRNALys + diphosphate
-
best noncognate amino acid substrate, 1600fold lower activity than with L-lysine, addition of 2 mM L-lysine abolishes the reaction
-
r
ATP + L-lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
-
-
-
?
ATP + L-lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
2-step reaction, the first step comprises the activation of the amino acid to form an enzyme-bound aminoacyl adenylate, the second step involves binding of this complex by tRNA, whose 3'-end is esterified with the aminoacyl-moiety followed by release of the resulting aminoacyl-tRNA, aminoacylation can be performed in absence of the tRNA
-
?
ATP + L-lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
cognate amino acid, best substrate, two-step reaction mechanism, limited selectivity in the aminoacylation reaction due to inefficient editing of some amino acids, e.g. Met, Leu, Cys, Ala, Thr, by pre-transfer mechanism or the absence of post-transfer editing of other amino acids e.g. Arg, Ser, a small fraction of lysine is converted to lysine lactam
-
r
ATP + L-lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
tRNA substrate from Borrelia burgdorferi or Escherichia coli, predicted secondary structure of tRNAlys from Escherichia coli
-
?
ATP + lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
-
-
-
?
ATP + lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
the enzymes major function is to provide Lys-tRNALys fpr protein biosynthesis
-
r
additional information
?
-
-
lysine-dependent ATP-diphosphate exchange reaction: lysine + ATP + enzyme/lysine-AMP-enzyme + diphosphate
-
-
?
additional information
?
-
-
synthesis of mixed dinucleoside 5',5'''-P1,P4-triphosphates or 5',5''''-P1,P4-tetraphosphates resulting from the reaction of lysyl adenylate with a variety of ribonucleotide 5'-triphosphates, ribonucleotide 5'-diphosphates, deoxyribonucleotide 5'-diphosphates or deoxyribonucleotide 5'-triphosphates
-
-
?
additional information
?
-
-
L-lysine-dependent synthesis of 5',5'-diadenosine tetraphosphate (Ap4A)
-
-
?
additional information
?
-
-
substrate recognition specificity, the enzyme also performs the ATP-diphosphate exchange reaction, class II lysine-tRNA synthetases recognize the same elements in tRNALys as their class I counterparts, namely the discriminator base N73 and the anticodon, but vary in the recognition of the G2.U71 wobble pair of spirochete tRNALys, which acts as a determinant for class II enzymes, but not for class I enzymes
-
?
additional information
?
-
-
the enzyme possesses an efficient pre-transfer editing mechanism which prevents misacylation of tRNALys with ornithine, which results in cyclization to ornithine lactam
-
?
additional information
?
-
-
the enzyme also converts ATP to diadenosine tri- and tetraphosphates in the presence of L-lysine/Mg2+/Zn2+
-
-
?
additional information
?
-
LysU-based preparation of potentially important ApnA analogues, overview. Dimeric LysU has dual diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) and diadenosine-5',5'''-P1,P3-triphosphate (Ap3A) synthase activities. Syntheses of both take place through the formation of a lysyl-adenylate 1 intermediate from ATP and L-lysine. Thereafter, the terminal phosphate of a second nucleotide substrate combines with the enzyme-bound lysyl-adenylate, thereby liberating free L-lysine and generating either Ap4A or Ap3A depending upon the identity of the second nucleotide substrate. The first step involving lysyladenylate intermediate formation is highly specific but reversible. Therefore inorganic diphosphatase-mediated controlled hydrolysis of diphosphate is required in order to prevent the back-reaction taking place, and thereby essentially rendering this first step committed. Fortunately, the second product formation step is highly promiscuous and a wide variety of nucleotide di-, tri-, and tetraphosphates are acceptable as second nucleotide substrates. This promiscuity also extends to inorganic phosphate and to tripolyphosphate. Surface mechanism of LysU catalyzed Ap4A and Ap3A synthase activities, reaction scheme and mechanism, overview. Bulkier putative diphosphate analogue substrates preclude molecular recognition and binding by LysU, hence preventing their use as bona fide LysU substrates able to couple to the ATP derived lysyl adenylate 1 intermediate with LysU assistance. Synthesis of analogues beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)2-beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)2-beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)-beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)-beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, alpha,beta-methylene 5'-P1,P3-bis(5'-adenosyl) triphosphate, alpha,beta-methylene-guanosine 5'-P1-triphospho-P3-5''-adenosine, beta,gamma-methylene-P1,P5-bis(5'-adenosyl) pentaphosphate, beta,gamma-imido-adenosine 5'-P1-pentaphospho-P5-5''-uridine, beta,gamma-methylene-adenosine 5'-P1-tretraphospho-P4-5''-guanosine, beta,gamma-imido-adenosine 5'-P1-tretraphospho-P4-5''-guanosine, and beta,gamma-delta,epsilon-dimethylene-P1,P6-bis(5'-adenosyl) hexaphosphate
-
-
?
additional information
?
-
-
LysU-based preparation of potentially important ApnA analogues, overview. Dimeric LysU has dual diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) and diadenosine-5',5'''-P1,P3-triphosphate (Ap3A) synthase activities. Syntheses of both take place through the formation of a lysyl-adenylate 1 intermediate from ATP and L-lysine. Thereafter, the terminal phosphate of a second nucleotide substrate combines with the enzyme-bound lysyl-adenylate, thereby liberating free L-lysine and generating either Ap4A or Ap3A depending upon the identity of the second nucleotide substrate. The first step involving lysyladenylate intermediate formation is highly specific but reversible. Therefore inorganic diphosphatase-mediated controlled hydrolysis of diphosphate is required in order to prevent the back-reaction taking place, and thereby essentially rendering this first step committed. Fortunately, the second product formation step is highly promiscuous and a wide variety of nucleotide di-, tri-, and tetraphosphates are acceptable as second nucleotide substrates. This promiscuity also extends to inorganic phosphate and to tripolyphosphate. Surface mechanism of LysU catalyzed Ap4A and Ap3A synthase activities, reaction scheme and mechanism, overview. Bulkier putative diphosphate analogue substrates preclude molecular recognition and binding by LysU, hence preventing their use as bona fide LysU substrates able to couple to the ATP derived lysyl adenylate 1 intermediate with LysU assistance. Synthesis of analogues beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)2-beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)2-beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)-beta,gamma-methylene-P1,P4-bis(5'-adenosyl) tetraphosphate, (open-ring-ribosyl)-beta,gamma-imido-P1,P4-bis(5'-adenosyl) tetraphosphate, alpha,beta-methylene 5'-P1,P3-bis(5'-adenosyl) triphosphate, alpha,beta-methylene-guanosine 5'-P1-triphospho-P3-5''-adenosine, beta,gamma-methylene-P1,P5-bis(5'-adenosyl) pentaphosphate, beta,gamma-imido-adenosine 5'-P1-pentaphospho-P5-5''-uridine, beta,gamma-methylene-adenosine 5'-P1-tretraphospho-P4-5''-guanosine, beta,gamma-imido-adenosine 5'-P1-tretraphospho-P4-5''-guanosine, and beta,gamma-delta,epsilon-dimethylene-P1,P6-bis(5'-adenosyl) hexaphosphate
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + L-lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
-
?
ATP + lysine + tRNALys
AMP + L-lysyl-tRNALys + diphosphate
-
the enzymes major function is to provide Lys-tRNALys fpr protein biosynthesis
-
r
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
NH4+
-
synthesis of lysyl-tRNA from lysyl-AMP-enzyme is dependent on NH4+ or other monovalent cations
Zn2+
Mg2+
-
formation of Lys-tRNALys from lysine, tRNA and ATP-enzyme complex or ATP is dependent on Mg2+
Mg2+
-
requires three Mg2+ in the activation reaction (including the one in MgATP). Both MgPPi and Mg2PPi, participate in the pyrophosphorolysis of the aminoacyl adenylate
Zn2+
-
0.06 mM, 50% inhibition of aminoacylation, stimulation of synthesis of mixed dinucleoside 5',5'''-P1,P4-tri- or 5',5''''-P1,P4-tetraphosphates resulting from the reaction of lysyl adenylate with a variety of ribo- (or deoxyribo-)nucleotides 5'-tri-(or 5'-di-)phosphates
Zn2+
-
stimulation of diadenosine 5',5'''-P1,P4-tetraphosphate synthetase activity
Zn2+
-
required, the wild-type enzyme has a weaker glycerol kinase-like capability in the absence of Zn2+ and is dominated by diadenosine tri- and tetraphosphate synthesis in its presence
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Mg2+
-
synthesis of lysyl-tRNA from lysyl-AMP-enzyme inhibited, formation of Lys-tRNALys from lysine, tRNA and ATP-enzyme complex or ATP is dependent on Mg2+
NH4+
-
formation of Lys-tRNALys from lysine, tRNA and ATP-enzyme complex or ATP
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.055
Escherichia coli wild type tRNALys
-
recombinant enzyme, pH 7.2, 37°C
-
0.00067
L-arginyl-tRNALys
-
deacylation reaction, pH 7.4, 37°C, in absence or presence of 50 mM DTT
0.00056
L-methionyl-tRNALys
-
deacylation reaction, pH 7.4, 37°C, in absence or presence of 50 mM DTT
-
0.00076
L-threonyl-tRNALys
-
deacylation reaction, pH 7.4, 37°C, in absence or presence of 50 mM DTT
-
0.00023
L-lysyl-tRNALys
-
deacylation reaction, pH 7.4, 37°C, in absence of DTT
0.0088
L-lysyl-tRNALys
-
deacylation reaction, pH 7.4, 37°C, in presence of 50 mM DTT
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00001
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme E240Q
0.000011
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme E428Q
0.000023
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme E278Q
0.000028
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, wild-type enzyme
0.000029
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme E240D
0.000039
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme N424D
0.000041
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme E278D
0.00005
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme F426W
0.000055
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme N424Q
0.000059
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme G216A
0.000061
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme E428D
0.00022
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme Y280S
0.00027
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme Y280F
0.0013
5'-O-[N-(L-lysyl)sulfamoyl]adenosine
-
pH 7.2, 37°C, mutant enzyme F426H
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
editing and aminoacylation reaction with several amino acids, up to 500fold variations in catalytic efficiencies
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
LysU is useful as a tool for highly controlled phosphate-phosphate bond formation between nucleotides, avoiding the need for complex protecting group chemistries. Resulting high yielding tandem LysU-based biosynthetic-synthetic/synthetic-biosynthetic strategies emerge for the preparation of varieties of ApnA analogues directly from inexpensive natural nucleotides and nucleosides. Analogues so formed make a useful small library with which to probe ApnA activities in vitro and in vivo leading to the discovery of potentially potent biopharmaceuticals active against chronic pain and other chronic, high-burden disease states
additional information
-
LysU is useful as a tool for highly controlled phosphate-phosphate bond formation between nucleotides, avoiding the need for complex protecting group chemistries. Resulting high yielding tandem LysU-based biosynthetic-synthetic/synthetic-biosynthetic strategies emerge for the preparation of varieties of ApnA analogues directly from inexpensive natural nucleotides and nucleosides. Analogues so formed make a useful small library with which to probe ApnA activities in vitro and in vivo leading to the discovery of potentially potent biopharmaceuticals active against chronic pain and other chronic, high-burden disease states
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
unliganded enzyme and in complex with L-lysine, vapor diffusion hanging drop method, protein solution: 20 mg/ml protein, 20 mM Tris-HCl, pH 8.0, 10 mM 2-mercaptoethanol, 10 mM MgCl2, plus equal volume of reservoir solution: 0.1 M HEPES, pH 7.5, 46-50% saturated ammonium sulfate solution, 2-4% polyethylene glycol 400, 15-20% glycerol, 4-18°C, X-ray difraction structure determination at 2.7 A resolution, and analysis
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E240D
-
1.2fold increase in Km-value for Lys, 21fold decrease in turnover number for Lys, Km-value for ATP is nearly identical to wild-type value, 24fold decrease in turnover number for ATP
E240Q
-
1.4fold increase in Km-value for Lys, 207fold decrease in turnover number for Lys, 1.2fold increase in Km-value for ATP, 261fold decrease in turnover number for ATP
E246D
-
the mutant shows more than 50% loss in catalytic efficiency compared to the wild type enzyme
E246R
-
the mutant shows more than 50% loss in catalytic efficiency compared to the wild type enzyme
E264A
-
the mutant shows more than 90% loss in catalytic efficiency compared to the wild type enzyme
E264K
-
the mutant catalyses the production of glycerol-3-phosphate, powered by ATP turnover to ADP but shows little formation of diadenosine tri- and tetraphosphates under normal conditions (additional Zn2+/L-lysine/Mg2+)
E264N
-
the mutant catalyses the production of glycerol-3-phosphate, powered by ATP turnover to ADP but shows little formation of diadenosine tri- and tetraphosphates under normal conditions (additional Zn2+/L-lysine/Mg2+)
E264Q
-
the mutant catalyses the production of glycerol-3-phosphate, powered by ATP turnover to ADP but shows little formation of diadenosine tri- and tetraphosphates under normal conditions (additional Zn2+/L-lysine/Mg2+)
E273A
-
the mutant shows more than 90% loss in catalytic efficiency compared to the wild type enzyme
E278D
-
98fold increase in Km-value for Lys, 11fold decrease in turnover number for Lys, 1.3fold increase in Km-value for ATP, 63fold decrease in turnover number for ATP
E278Q
-
1.9fold decrease in Km-value for Lys, 120fold decrease in turnover number for Lys, 1.5fold decrease in Km-value for ATP, 200fold decrease in turnover number for ATP
E414A
-
the mutant shows more than 90% loss in catalytic efficiency compared to the wild type enzyme
E421A
-
the mutant shows more than 50% loss in catalytic efficiency compared to the wild type enzyme
E428D
-
8.5fold increase in Km-value for Lys, 1.8fold decrease in turnover number for Lys, 1.7fold increase in Km-value for ATP, 3.4fold decrease in turnover number for ATP
E428Q
-
2fold decrease in Km-value for Lys, 9fold decrease in turnover number for Lys, 1.5fold decrease in Km-value for ATP, 200fold decrease in turnover number for ATP
F261A
-
the mutant shows more than 50% loss in catalytic efficiency compared to the wild type enzyme
F274A
-
the mutant shows more than 90% loss in catalytic efficiency compared to the wild type enzyme
F426H
-
78fold increase in Km-value for Lys, 1.3fold decrease in turnover number for Lys, 1.6fold increase in Km-value for ATP, 5fold decrease in turnover number for ATP
F426W
-
6.2fold increase in Km-value for Lys, 3fold decrease in turnover number for Lys, 9.2fold increase in Km-value for ATP, 1.4fold decrease in turnover number for ATP
G216A
-
75fold increase in Km-value for Lys, 1.9fold increase in turnover number for Lys, 16fold increase in Km-value for ATP, 4.25fold decrease in turnover number for ATP
G265A
-
the mutant shows more than 90% loss in catalytic efficiency compared to the wild type enzyme
H270A
-
the mutant catalyses the production of glycerol-3-phosphate, powered by ATP turnover to ADP but shows little formation of diadenosine tri- and tetraphosphates under normal conditions (additional Zn2+/L-lysine/Mg2+)
I266A
-
the mutant shows more than 50% loss in catalytic efficiency compared to the wild type enzyme
N424D
-
2fold increase in Km-value for Lys, 1.1fold decrease in turnover number for Lys, 1.8fold increase in Km-value for ATP, 3.4fold decrease in turnover number for ATP
N424Q
-
20fold increase in Km-value for Lys, 2.8fold decrease in turnover number for Lys, 69fold increase in Km-value for ATP, 1.5fold decrease in turnover number for ATP
P272A
-
the mutant shows more than 90% loss in catalytic efficiency compared to the wild type enzyme
R480A
-
the mutant shows more than 50% loss in catalytic efficiency compared to the wild type enzyme
S267A
-
the mutant shows more than 50% loss in catalytic efficiency compared to the wild type enzyme
Y280F
-
9.2fold increase in Km-value for Lys, 58fold decrease in turnover number for Lys, 5.2fold increase in Km-value for ATP, 20fold decrease in turnover number for ATP
Y280S
-
44fold increase in Km-value for Lys, 1.1fold decrease in turnover number for Lys, 3.9fold increase in Km-value for ATP, 9.7fold decrease in turnover number for ATP
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, Q Sepharose column chromatography, and Sephacryl S300 gel filtration
-
recombinant His-tagged protein from stran BL21(DE3), functional complementation of a class II-enzyme-deficient Escherichia coli mutant
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug development
LysU is also useful as a tool for highly controlled phosphate-phosphate bond formation between nucleotides, avoiding the need for complex protecting group chemistries. Resulting high yielding tandem LysU-based biosynthetic-synthetic/synthetic-biosynthetic strategies emerge for the preparation of varieties of ApnA analogues directly from inexpensive natural nucleotides and nucleosides. Analogues so formed make a useful small library with which to probe ApnA activities in vitro and in vivo leading to the discovery of potentially potent biopharmaceuticals active against chronic pain and other chronic, high-burden disease states
synthesis
recombinant Escherichia coli lysyl-tRNA synthase (LysU) has been previously utilised in the production of stabile, synthetic diadenosine polyphosphate (ApnA) analogues. LysU is also useful as a tool for highly controlled phosphate-phosphate bond formation between nucleotides, avoiding the need for complex protecting group chemistries. Resulting high yielding tandem LysU-based biosynthetic-synthetic/synthetic-biosynthetic strategies emerge for the preparation of varieties of ApnA analogues directly from inexpensive natural nucleotides and nucleosides. Analogues so formed make a useful small library with which to probe ApnA activities in vitro and in vivo leading to the discovery of potentially potent biopharmaceuticals active against chronic pain and other chronic, high-burden disease states
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Airas, R.K.
Differences in the magnesium dependences of the class I and class II aminoacyl-tRNA synthetases from Escherichia coli
Eur. J. Biochem.
240
223-231
1996
Escherichia coli
Onesti, S.; Miller, A.D.; Brick, P.
The crystal structure of the lysyl-tRNA synthetase from Escherichia coli
Structure
15
163-176
1995
Escherichia coli
Hele, P.; Barber, R.
Lysyl tRNA synthetase of Escherichia coli. Formation and reactions of ATP-enzyme and lysyl-AMP-enzyme complexes
Biochim. Biophys. Acta
258
319-331
1972
Escherichia coli, Escherichia coli B / ATCC 11303
Kisselev, L.L.; Baturina, I.D.
Two enzymatically active forms of lysyl-tRNA synthetase from E. coli B
FEBS Lett.
22
231-234
1972
Escherichia coli, Escherichia coli B / ATCC 11303
Baturina, I.D.; Gnutchev, N.V.; Khomutov, R.M.; Kisselev, L.L.
Substrate specificity of lysyl-tRNA synthetase from E. coli B
FEBS Lett.
22
235-237
1972
Escherichia coli, Escherichia coli B / ATCC 11303
Dittgen, R.M.; Leberman, R.
Multiple forms of lysyl-tRNA synthetase from Escherichia coli
Hoppe-Seyler's Z. Physiol. Chem.
357
543-551
1976
Escherichia coli, Escherichia coli MRE 600
Plateau, P.; Blanquet, S.
Zinc-dependent synthesis of various dinucleoside 5', 5'''-P1, P3-tri or 5', 5'''-P1,P4-tetraphosphates by Escherichia coli lysyl-tRNA synthetase
Biochemistry
21
5273-5279
1982
Escherichia coli
Jakubowski, H.
Misacylation of tRNALys with noncognate amino acids by lysyl-tRNA synthetase
Biochemistry
38
8088-8093
1999
Escherichia coli
Onesti, S.; Desogus, G.; Brevet, A.; Chen, J.; Plateau, P.; Blanquet, S.; Brick, P.
Structural studies of lysyl-tRNA synthetase: conformational changes induced by substrate binding
Biochemistry
39
12853-12861
2000
Escherichia coli (P0A8N3), Escherichia coli
Levengood, J.D.; Ataide, S.F.; Roy, H.; Ibba, M.
Divergence in non-cognate amino acid recognition between class I and class II lysyl-tRNA synthetases
J. Biol. Chem.
279
17707-17714
2004
Borreliella burgdorferi, Escherichia coli
Ibba, M.; Losey, H.C.; Kawarabayasi, Y.; Kikuchi, H.; Bunjun, S.; Soll, D.
Substrate recognition by class I lysyl-tRNA synthetases: a molecular basis for gene displacement
Proc. Natl. Acad. Sci. USA
96
418-423
1999
Aeropyrum pernix (Q9YFT9), Borreliella burgdorferi, Escherichia coli, Methanococcus maripaludis
Ataide, S.F.; Ibba, M.
Discrimination of cognate and noncognate substrates at the active site of class II lysyl-tRNA synthetase
Biochemistry
43
11836-11841
2004
Escherichia coli
Wright, M.; Boonyalai, N.; Tanner, J.A.; Hindley, A.D.; Miller, A.D.
The duality of LysU, a catalyst for both Ap4A and Ap3A formation
FEBS J.
273
3534-3544
2006
Escherichia coli
Fukunaga, J.; Yokogawa, T.; Ohno, S.; Nishikawa, K.
Misacylation of yeast amber suppressor tRNA(Tyr) by E. coli lysyl-tRNA synthetase and its effective repression by genetic engineering of the tRNA sequence
J. Biochem.
139
689-696
2006
Escherichia coli
Chou, T.F.; Wagner, C.R.
Lysyl-tRNA synthetase-generated lysyl-adenylate is a substrate for histidine triad nucleotide binding proteins
J. Biol. Chem.
282
4719-4727
2007
Escherichia coli, Homo sapiens
Fukunaga, J.; Ohno, S.; Nishikawa, K.; Yokogawa, T.
A base pair at the bottom of the anticodon stem is reciprocally preferred for discrimination of cognate tRNAs by Escherichia coli lysyl- and glutaminyl-tRNA synthetases
Nucleic Acids Res.
34
3181-3188
2006
Escherichia coli
Chen, X.; Boonyalai, N.; Lau, C.; Thipayang, S.; Xu, Y.; Wright, M.; Miller, A.D.
Multiple catalytic activities of Escherichia coli lysyl-tRNA synthetase (LysU) are dissected by site-directed mutagenesis
FEBS J.
280
102-114
2013
Escherichia coli
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