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Enzyme Nomenclature
Information on EC 6.2.1.68 - L-glutamate---[L-glutamyl-carrier protein] ligase
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EC Tree
6.2.1.68 L-glutamate---[L-glutamyl-carrier protein] ligaseIUBMB Comments
The adenylation domain of the enzyme catalyses the activation of L-glutamate to (L-glutamyl)adenylate, followed by the transfer of the activated compound to the free thiol of a phosphopantetheine arm of a peptidyl-carrier protein domain. The peptidyl-carrier protein domain may be part of the same protein, or of a different protein. This activity is often found as part of a larger non-ribosomal peptide synthase.
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The expected taxonomic range for this enzyme is: Pseudomonas aeruginosa
Reaction Schemes
+
+
=
+
+
+
+
holo-[L-glutamyl-carrier protein]
=
+
+
L-glutamyl-[L-glutamyl-carrier protein]
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
ambE
non-ribosomal peptide synthetase
additional information
ambE
-
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] = AMP + L-glutamyl-[L-glutamyl-carrier protein]
(1b)
-
-
-
ATP + L-glutamate + holo-[L-glutamyl-carrier protein] = AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
overall reaction
-
-
-
ATP + L-glutamate = diphosphate + (L-glutamyl)adenylate
(1a)
-
-
-
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SYSTEMATIC NAME
IUBMB Comments
L-glutamate:[L-glutamyl-carrier protein] ligase (AMP-forming)
The adenylation domain of the enzyme catalyses the activation of L-glutamate to (L-glutamyl)adenylate, followed by the transfer of the activated compound to the free thiol of a phosphopantetheine arm of a peptidyl-carrier protein domain. The peptidyl-carrier protein domain may be part of the same protein, or of a different protein. This activity is often found as part of a larger non-ribosomal peptide synthase.
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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
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
additional information
?
-
identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview
-
-
-
additional information
?
-
identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview
-
-
-
additional information
?
-
identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview
-
-
-
additional information
?
-
identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview
-
-
-
additional information
?
-
identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview
-
-
-
additional information
?
-
identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview
-
-
-
additional information
?
-
identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview
-
-
-
additional information
?
-
identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview
-
-
-
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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
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
(L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate
diphosphate + (L-glutamyl)adenylate
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
-
-
ir
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4'-phosphopantetheine
phosphopantetheinylation of recombinant apoenzyme AmbE by a purified promiscuous phosphopantetheinyl transferase Sfp at pH 7.5, 28°C
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
physiological function
additional information
metabolism
Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide
metabolism
-
Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide
-
metabolism
-
Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide
-
metabolism
-
Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide
-
metabolism
-
Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide
-
metabolism
-
Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide
-
metabolism
-
Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide
-
metabolism
-
Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide
-
physiological function
the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview
physiological function
-
the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview
-
physiological function
-
the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview
-
physiological function
-
the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview
-
physiological function
-
the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview
-
physiological function
-
the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview
-
physiological function
-
the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview
-
physiological function
-
the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview
-
additional information
enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs)
additional information
-
enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs)
-
additional information
-
enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs)
-
additional information
-
enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs)
-
additional information
-
enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs)
-
additional information
-
enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs)
-
additional information
-
enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs)
-
additional information
-
enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs)
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). AMBE_PSEAE
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
2124
0
229044
Swiss-Prot
-
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview
additional information
-
modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview
-
additional information
-
modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview
-
additional information
-
modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview
-
additional information
-
modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview
-
additional information
-
modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview
-
additional information
-
modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview
-
additional information
-
modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview
-
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D644A
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
D644A/K1230T/S1985A
site-directed mutagenesis, the mutations abolish loading of L-Glu completely
K1230T
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
S1286A
site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely
S1819A
site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu
S1958A
site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu
D644A
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
K1230T
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
S1286A
-
site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely
-
S1819A
-
site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu
-
S1958A
-
site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu
-
D644A
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
K1230T
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
S1286A
-
site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely
-
S1819A
-
site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu
-
S1958A
-
site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu
-
D644A
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
K1230T
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
S1286A
-
site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely
-
S1819A
-
site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu
-
S1958A
-
site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu
-
D644A
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
K1230T
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
S1286A
-
site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely
-
S1819A
-
site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu
-
S1958A
-
site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu
-
D644A
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
K1230T
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
S1286A
-
site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely
-
S1819A
-
site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu
-
S1958A
-
site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu
-
D644A
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
K1230T
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
S1286A
-
site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely
-
S1819A
-
site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu
-
S1958A
-
site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu
-
D644A
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
K1230T
-
site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely
-
S1286A
-
site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely
-
S1819A
-
site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu
-
S1958A
-
site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu
-
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)/pLys by nickel affinity chromatography
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene ambE, encoded in the amb gene cluster, genetic organization, recombinant overexpression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)/pLys
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Murcia, N.; Lee, X.; Waridel, P.; Maspoli, A.; Imker, H.; Chai, T.; Walsh, C.; Reimmann, C.
The Pseudomonas aeruginosa antimetabolite L-2-amino-4-methoxy-trans-3-butenoic acid (AMB) is made from glutamate and two alanine residues via a thiotemplate-linked tripeptide precursor
Front. Microbiol.
6
170
2015
Pseudomonas aeruginosa (Q9I1H3), Pseudomonas aeruginosa ATCC 15692 (Q9I1H3), Pseudomonas aeruginosa 1C (Q9I1H3), Pseudomonas aeruginosa PRS 101 (Q9I1H3), Pseudomonas aeruginosa DSM 22644 (Q9I1H3), Pseudomonas aeruginosa CIP 104116 (Q9I1H3), Pseudomonas aeruginosa LMG 12228 (Q9I1H3), Pseudomonas aeruginosa JCM 14847 (Q9I1H3)
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