BRENDA - Enzyme Database
show all sequences of 6.3.5.6

The heterotrimeric Thermus thermophilus Asp-tRNA(Asn) amidotransferase can also generate Gln-tRNA(Gln)

Becker, H.D.; Min, B.; Jacobi, C.; Raczniak, G.; Pelaschier, J.; Roy, H.; Klein, S.; Kern, D.; Sll, D.; FEBS Lett. 476, 140-144 (2000)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
canonical set of amidotransferase genes gatA, gatB and gatC is cloned and sequenced, the holoenzyme is overexpessed in Escherichia coli and exhibits both Asp-tRNAAsn and Glu-tRNAGln transamidation activities, the gatA, gatB and gatC genes are dispersed in the genome and encode polypeptides of 472, 470 and 90 amino acids, removal of the 44 carboxy-terminal amino acids of the GatA subunit only inhibits the Asp-AdT activity, but not the Glu-AdT activity
Thermus thermophilus
Engineering
Amino acid exchange
Commentary
Organism
additional information
GatDDELTA44 mutant AdT possesses a GatA subunit deprived of its natural 44 C-terminal amino acids and shows unaltered Glu-AdT activity, but lacks any detectable Asp-AdT activity, deletion of one base in comparison with the HB27 genome causes a frameshift mutation
Thermus thermophilus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
Asp-tRNAAsn + ?
Thermus thermophilus
in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity
Asn-tRNAAsn + ?
-
Thermus thermophilus
?
Asp-tRNAAsn + ?
Deinococcus radiodurans
in vivo only Asn-tRNAAsn formation occurs
Asn-tRNAAsn + ?
-
Deinococcus radiodurans
?
Asp-tRNAAsn + ?
Thermus thermophilus
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
Asn-tRNAAsn + ?
-
Thermus thermophilus
?
Asp-tRNAAsn + ?
Deinococcus radiodurans
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
Asn-tRNAAsn + ?
-
Deinococcus radiodurans
?
Asp-tRNAAsn + ?
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity
Asn-tRNAAsn + ?
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
Asp-tRNAAsn + ?
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
Asn-tRNAAsn + ?
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Bacillus subtilis
-
-
-
Deinococcus radiodurans
-
-
-
Thermus thermophilus
-
and strain HB27 / ATCC BAA-163 / DSM 7039
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
-
and strain HB27 / ATCC BAA-163 / DSM 7039
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Asp-tRNAAsn + ?
in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity
440090
Thermus thermophilus
Asn-tRNAAsn + ?
-
440090
Thermus thermophilus
?
Asp-tRNAAsn + ?
in vivo only Asn-tRNAAsn formation occurs
440090
Deinococcus radiodurans
Asn-tRNAAsn + ?
-
440090
Deinococcus radiodurans
?
Asp-tRNAAsn + ?
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
440090
Thermus thermophilus
Asn-tRNAAsn + ?
-
440090
Thermus thermophilus
?
Asp-tRNAAsn + ?
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
440090
Deinococcus radiodurans
Asn-tRNAAsn + ?
-
440090
Deinococcus radiodurans
?
Asp-tRNAAsn + ?
in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
Asn-tRNAAsn + ?
-
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
Asp-tRNAAsn + ?
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
Asn-tRNAAsn + ?
-
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
ATP + Asp-tRNAAsn + glutamine
-
440090
Bacillus subtilis
ADP + phosphate + Asn-tRNAAsn + glutamate
-
440090
Bacillus subtilis
?
ATP + Asp-tRNAAsn + glutamine
-
440090
Deinococcus radiodurans
ADP + phosphate + Asn-tRNAAsn + glutamate
-
440090
Deinococcus radiodurans
?
ATP + Asp-tRNAAsn + glutamine
no restricted substrate specificity of tRNA-dependent AdT: dual-specific Asp/Glu-AdT, enzyme catalyzes transamidation of Asp-tRNAAsn, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, GatA subunit can discriminate between Asp-tRNAAsn and Glu-tRNAGln and therefore is implicated in binding of the tRNA
440090
Thermus thermophilus
ADP + phosphate + Asn-tRNAAsn + glutamate
forms Asn-tRNAAsn
440090
Thermus thermophilus
?
ATP + Asp-tRNAAsn + glutamine
no restricted substrate specificity of tRNA-dependent AdT: dual-specific Asp/Glu-AdT, enzyme catalyzes transamidation of Asp-tRNAAsn, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, GatA subunit can discriminate between Asp-tRNAAsn and Glu-tRNAGln and therefore is implicated in binding of the tRNA
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
ADP + phosphate + Asn-tRNAAsn + glutamate
forms Asn-tRNAAsn
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
ATP + Glu-tRNAGln + glutamine
in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Gln-tRNAGln formation
440090
Bacillus subtilis
ADP + Gln-tRNAGln + glutamate
-
440090
Bacillus subtilis
?
ATP + Glu-tRNAGln + glutamine
in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation
440090
Thermus thermophilus
ADP + Gln-tRNAGln + glutamate
-
440090
Thermus thermophilus
?
ATP + Glu-tRNAGln + glutamine
in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation
440090
Deinococcus radiodurans
ADP + Gln-tRNAGln + glutamate
-
440090
Deinococcus radiodurans
?
ATP + Glu-tRNAGln + glutamine
in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
ADP + Gln-tRNAGln + glutamate
-
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
additional information
-
440090
Deinococcus radiodurans
?
-
-
-
-
additional information
cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases
440090
Thermus thermophilus
?
-
-
-
-
additional information
cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
heterodimer
Asp-AdT is active as a GatAB heterodimer
Thermus thermophilus
heterotrimer
possesses the three canonical gatA, gatB and gatC set of genes encoding heterotrimeric AdT: GatABC, GatA and GatB are more tightly bound to each other than to GatC, GatA subunit has conserved Gln-436
Thermus thermophilus
heterotrimer
-
Bacillus subtilis
heterotrimer
-
Deinococcus radiodurans
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
assay at, suboptimal temperature for the thermophilic enzyme
Thermus thermophilus
Temperature Stability [C]
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
additional information
-
thermophilic enzyme
Thermus thermophilus
Cloned(Commentary) (protein specific)
Commentary
Organism
canonical set of amidotransferase genes gatA, gatB and gatC is cloned and sequenced, the holoenzyme is overexpessed in Escherichia coli and exhibits both Asp-tRNAAsn and Glu-tRNAGln transamidation activities, the gatA, gatB and gatC genes are dispersed in the genome and encode polypeptides of 472, 470 and 90 amino acids, removal of the 44 carboxy-terminal amino acids of the GatA subunit only inhibits the Asp-AdT activity, but not the Glu-AdT activity
Thermus thermophilus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
GatDDELTA44 mutant AdT possesses a GatA subunit deprived of its natural 44 C-terminal amino acids and shows unaltered Glu-AdT activity, but lacks any detectable Asp-AdT activity, deletion of one base in comparison with the HB27 genome causes a frameshift mutation
Thermus thermophilus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
Asp-tRNAAsn + ?
Thermus thermophilus
in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity
Asn-tRNAAsn + ?
-
Thermus thermophilus
?
Asp-tRNAAsn + ?
Deinococcus radiodurans
in vivo only Asn-tRNAAsn formation occurs
Asn-tRNAAsn + ?
-
Deinococcus radiodurans
?
Asp-tRNAAsn + ?
Thermus thermophilus
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
Asn-tRNAAsn + ?
-
Thermus thermophilus
?
Asp-tRNAAsn + ?
Deinococcus radiodurans
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
Asn-tRNAAsn + ?
-
Deinococcus radiodurans
?
Asp-tRNAAsn + ?
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity
Asn-tRNAAsn + ?
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
Asp-tRNAAsn + ?
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
Asn-tRNAAsn + ?
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Asp-tRNAAsn + ?
in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity
440090
Thermus thermophilus
Asn-tRNAAsn + ?
-
440090
Thermus thermophilus
?
Asp-tRNAAsn + ?
in vivo only Asn-tRNAAsn formation occurs
440090
Deinococcus radiodurans
Asn-tRNAAsn + ?
-
440090
Deinococcus radiodurans
?
Asp-tRNAAsn + ?
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
440090
Thermus thermophilus
Asn-tRNAAsn + ?
-
440090
Thermus thermophilus
?
Asp-tRNAAsn + ?
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
440090
Deinococcus radiodurans
Asn-tRNAAsn + ?
-
440090
Deinococcus radiodurans
?
Asp-tRNAAsn + ?
in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
Asn-tRNAAsn + ?
-
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
Asp-tRNAAsn + ?
enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
Asn-tRNAAsn + ?
-
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
ATP + Asp-tRNAAsn + glutamine
-
440090
Bacillus subtilis
ADP + phosphate + Asn-tRNAAsn + glutamate
-
440090
Bacillus subtilis
?
ATP + Asp-tRNAAsn + glutamine
-
440090
Deinococcus radiodurans
ADP + phosphate + Asn-tRNAAsn + glutamate
-
440090
Deinococcus radiodurans
?
ATP + Asp-tRNAAsn + glutamine
no restricted substrate specificity of tRNA-dependent AdT: dual-specific Asp/Glu-AdT, enzyme catalyzes transamidation of Asp-tRNAAsn, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, GatA subunit can discriminate between Asp-tRNAAsn and Glu-tRNAGln and therefore is implicated in binding of the tRNA
440090
Thermus thermophilus
ADP + phosphate + Asn-tRNAAsn + glutamate
forms Asn-tRNAAsn
440090
Thermus thermophilus
?
ATP + Asp-tRNAAsn + glutamine
no restricted substrate specificity of tRNA-dependent AdT: dual-specific Asp/Glu-AdT, enzyme catalyzes transamidation of Asp-tRNAAsn, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, GatA subunit can discriminate between Asp-tRNAAsn and Glu-tRNAGln and therefore is implicated in binding of the tRNA
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
ADP + phosphate + Asn-tRNAAsn + glutamate
forms Asn-tRNAAsn
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
ATP + Glu-tRNAGln + glutamine
in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Gln-tRNAGln formation
440090
Bacillus subtilis
ADP + Gln-tRNAGln + glutamate
-
440090
Bacillus subtilis
?
ATP + Glu-tRNAGln + glutamine
in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation
440090
Thermus thermophilus
ADP + Gln-tRNAGln + glutamate
-
440090
Thermus thermophilus
?
ATP + Glu-tRNAGln + glutamine
in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation
440090
Deinococcus radiodurans
ADP + Gln-tRNAGln + glutamate
-
440090
Deinococcus radiodurans
?
ATP + Glu-tRNAGln + glutamine
in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
ADP + Gln-tRNAGln + glutamate
-
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
additional information
-
440090
Deinococcus radiodurans
?
-
-
-
-
additional information
cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases
440090
Thermus thermophilus
?
-
-
-
-
additional information
cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases
440090
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
heterodimer
Asp-AdT is active as a GatAB heterodimer
Thermus thermophilus
heterotrimer
possesses the three canonical gatA, gatB and gatC set of genes encoding heterotrimeric AdT: GatABC, GatA and GatB are more tightly bound to each other than to GatC, GatA subunit has conserved Gln-436
Thermus thermophilus
heterotrimer
-
Bacillus subtilis
heterotrimer
-
Deinococcus radiodurans
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
assay at, suboptimal temperature for the thermophilic enzyme
Thermus thermophilus
Temperature Stability [C] (protein specific)
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
additional information
-
thermophilic enzyme
Thermus thermophilus
Other publictions for EC 6.3.5.6
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
746380
Vijayakumar
Soluble expression and purifi ...
Fasciola gigantica
Protein Expr. Purif.
143
9-13
2018
-
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1
-
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1
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1
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1
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1
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728743
Fuengfuloy
Overproduction of the N-termin ...
no activity in Helicobacter pylori
Protein Expr. Purif.
89
25-32
2013
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1
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728397
Fischer
The asparagine-transamidosome ...
Helicobacter pylori
Nucleic Acids Res.
40
4965-4976
2012
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-
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1
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1
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2
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1
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1
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1
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1
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728424
Yu
New WS9326A congeners from Str ...
Brugia malayi
Org. Lett.
14
4946-4949
2012
-
-
-
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4
-
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1
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2
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2
4
-
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715924
Crepin
A hybrid structural model of t ...
Brugia malayi
J. Mol. Biol.
405
1056-1069
2011
-
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1
1
1
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1
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1
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1
1
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716420
Yu
Tirandamycins from Streptomyce ...
Brugia malayi
Org. Lett.
13
2034-2037
2011
-
1
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1
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1
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