Information on EC 3.4.17.11 - glutamate carboxypeptidase

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

EC NUMBER
COMMENTARY hide
3.4.17.11
-
RECOMMENDED NAME
GeneOntology No.
glutamate carboxypeptidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
release of C-terminal glutamate residues from a wide range of N-acylating moieties, including peptidyl, aminoacyl, benzoyl, benzyloxycarbonyl, folyl and pteroyl groups
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
9074-87-7
not distinguishable from EC 3.4.19.9 in Chemical Abstracts
94894-32-3
Pseudomonas isoenzyme 2
94894-33-4
Pseudomonas isoenzyme 2 precursor
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
strain RS16, gene CPG2
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2-chloroethyl)-(2-(methylsulfoyloxyethyl)amino)aminobenzyl-L-glutamic acid + H2O
(2-chloroethyl)-(2-(methylsulfoyloxyethyl)amino)aminobenzoic acid + L-glutamate
show the reaction diagram
-
a prodrug
-
-
?
(2S,2'S)-2,2'-(pentane-1,5-diylbis[oxy[(11S,11aS)-11-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8,10(5H)-diyl]carbonyloxymethanediylbenzene-4,1-diylcarbamoylimino])dipentanedioic acid + 2 H2O
(11aS,11a'S)-8,8'-[pentane-1,5-diylbis(oxy)]bis(7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one) + ?
show the reaction diagram
-
-
-
-
?
(2S,2'S)-2,2'-(pentane-1,5-diylbis[oxy[(11S,11aS)-11-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8,10(5H)-diyl]carbonyloxymethanediylbenzene-4,1-diyloxycarbonylimino])dipentanedioic acid + 2 H2O
3-hydroxybenzyl 4-hydroxybenzyl (11S,11aS,11'S,11a'S)-8,8'-[pentane-1,5-diylbis(oxy)]bis(11-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-10(5H)-carboxylate) + 2 N-carboxy-L-glutamate
show the reaction diagram
-
-
-
-
?
(4-[bis(2-bromoethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid + H2O
4-[bis(2-bromoethyl)amino]-2,3,5-trifluorobenzoic acid + L-glutamate
show the reaction diagram
-
a trifluorinated prodrug
-
-
?
(4-[bis(2-bromoethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid + H2O
4-[bis(2-bromoethyl)amino]-3,5-difluorobenzoic acid + L-glutamate
show the reaction diagram
(4-[bis(2-chloroethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid + H2O
4-[bis(2-chloroethyl)amino]-2,3,5-trifluorobenzoic acid + L-glutamate
show the reaction diagram
-
a trifluorinated prodrug
-
-
?
(4-[bis(2-chloroethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid + H2O
4-[bis(2-chloroethyl)amino]-3,5-difluorobenzoic acid + L-glutamate
show the reaction diagram
-
a difluorinated prodrug
-
-
?
(4-[bis(2-iodoethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid + H2O
4-[bis(2-iodoethyl)amino]-2,3,5-trifluorobenzoic acid + L-glutamate
show the reaction diagram
-
a trifluorinated prodrug
-
-
?
(4-[bis(2-iodoethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid + H2O
4-[bis(2-iodoethyl)amino]-3,5-difluorobenzoic acid + L-glutamate
show the reaction diagram
(6R,S)-leucovorin + H2O
N-(6R,S)-5-formyl-5,6,7,8-tetrahydropteroic acid + L-glutamate
show the reaction diagram
2,4-dihydroxypteroyl-L-glutamic acid + H2O
2,4-dihydroxypteroate + L-Glu
show the reaction diagram
-
-
-
-
?
3,5-difluoro-4-[bis(2-iodoethyl)amino]benzoic acid + H2O
?
show the reaction diagram
3,5-difluoro-4-[bis(2-iodoethyl)amino]benzoyl-L-glutamic acid + H2O
?
show the reaction diagram
3,5-difluorobenzoyl-L-glutamate + H2O
3,5-difluorobenzoate + L-glutamate
show the reaction diagram
3,5-difluorobenzoyl-L-glutamic acid + H2O
3,5-difluorobenzoic acid + L-glutamate
show the reaction diagram
-
-
-
-
?
3-fluoro-4-[bis(2-chloroethyl)amino]benzoyl-L-glutamic acid + H2O
?
show the reaction diagram
4-([bis(2-iodoethyl)amino]-phenyl)oxycarbonyl-L-glutamic acid + H2O
4-[bis(2-iodoethyl)amino]-phenylcarbonate + L-glutamate
show the reaction diagram
4-amino-N10-methylpteroylaspartic acid + H2O
4-amino-N10-methylpteroate + Asp
show the reaction diagram
-
-
-
-
?
4-aminobenzoyl-L-glutamic acid + H2O
4-aminobenzoate + glutamate
show the reaction diagram
-
-
-
-
?
4-aminopteroylaspartic acid + H2O
4-aminopteroate + Asp
show the reaction diagram
-
-
-
-
?
4-aminopteroylglutamic acid + H2O
4-aminopteroate + Glu
show the reaction diagram
4-[(2-chloroethyl)(2-methylsulphonyloxyethyl)amino]benzoic acid + H2O
?
show the reaction diagram
-
cognate drug
-
-
?
4-[(2-chloroethyl)-(2-methylsulphonyloxyethyl)amino]benzoyl-L-glutamic acid + H2O
?
show the reaction diagram
-
prodrug activated by CPG2
-
-
?
4-[bis(2-iodoethyl)-amino]-phenyloxycarbonyl-L-glutamic acid + H2O
?
show the reaction diagram
-
prodrug activated by CPG2
-
-
?
5-formyltetrahydrofolic acid + H2O
5-formylpteroate + Glu
show the reaction diagram
5-methyltetrahydrofolate + H2O
5-methylpteroate + Glu
show the reaction diagram
7-hydroxy-methotrexate + H2O
7-hydroxy-4-amino-4-deoxy-N10-methylpteroic acid + L-glutamate
show the reaction diagram
-
-
-
-
?
alpha-L-glutamyl-L-glutamic acid + H2O
L-Glu + L-Glu
show the reaction diagram
-
-
-
-
?
benzoyl-L-glutamic acid + H2O
benzoate + glutamate
show the reaction diagram
-
-
-
-
?
di-tert-butyl 4-[[2'-[N,N-bis(2''-chloroethyl)amino]phenyl]carbamoyloxymethyl]phenyloxycarbonyl-L-glutamic acid + H2O
?
show the reaction diagram
diallyl 4-[N-[4'-bis(2''-chloroethyl)aminophenyl]-N-methylcarbamoyloxymethyl]phenylcarbamoyl-L-glutamic acid + H2O
?
show the reaction diagram
diallyl 4-[[2'-[N,N-bis(2''-chloroethyl)amino]phenyl]carbamoyloxymethyl]phenylcarbamoyl-L-glutamic acid + H2O
?
show the reaction diagram
diethylstilbestrol-glutamate + H2O
diethylstilbestrol + L-glutamate
show the reaction diagram
dihydrofolic acid + H2O
dihydropteroic acid + Glu
show the reaction diagram
-
-
-
-
?
folic acid + H2O
pteroic acid + glutamic acid
show the reaction diagram
gamma-L-glutamyl-L-glutamic acid + H2O
L-Glu + L-Glu
show the reaction diagram
-
-
-
-
?
glycyl-L-glutamic acid + H2O
glycine + glutamate
show the reaction diagram
-
-
-
-
?
L-tyrosyl-L-glutamic acid + H2O
tyrosine + glutamate
show the reaction diagram
-
-
-
-
?
methotrexate + H2O
4-amino-4-deoxy-N10-methylpteroic acid + L-glutamate
show the reaction diagram
methotrexate + H2O
4-[(2,4-diamino-6-(pteridinyl)methyl)-methylamino]-benzoic acid + L-glutamate
show the reaction diagram
methotrexate + H2O
?
show the reaction diagram
N-([4-[([[(11S,11aS)-11-hydroxy-7,8-dimethoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-10(5H)-yl]carbonyl]oxy)methyl]phenoxy]carbonyl)-L-glutamic acid + H2O
(11aS)-7,8-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one + ?
show the reaction diagram
-
-
-
-
?
N-([4-[([[(11S,11aS)-11-hydroxy-7,8-dimethoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-10(5H)-yl]carbonyl]oxy)methyl]phenyl]carbamoyl)-L-glutamic acid + H2O
? + L-glutamate
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-L-aspartic acid + H2O
benzyl carbonate + L-Asp
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-L-glutamic acid + H2O
benzyl carbonate + L-Glu
show the reaction diagram
-
no activity with D-glutamic acid
-
-
?
N-benzyloxycarbonyl-L-glutamine + H2O
benzyl carbonate + L-Gln
show the reaction diagram
-
-
-
-
?
pteroyl-gamma,gamma-triglutamate + H2O
?
show the reaction diagram
-
-
-
-
?
[4-[bis(2-chloroethyl)amino]-3,5-difluorobenzoyl]-L-glutamic acid + H2O
?
show the reaction diagram
-
prodrug activated by CPG2
-
-
?
[4-[bis(2-hydroxyethyl)amino]-3,5-difluorobenzoyl]-L-glutamic acid + H2O
?
show the reaction diagram
-
prodrug activated by CPG2
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(2S,2'S)-2,2'-(pentane-1,5-diylbis[oxy[(11S,11aS)-11-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8,10(5H)-diyl]carbonyloxymethanediylbenzene-4,1-diylcarbamoylimino])dipentanedioic acid + 2 H2O
(11aS,11a'S)-8,8'-[pentane-1,5-diylbis(oxy)]bis(7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one) + ?
show the reaction diagram
-
-
-
-
?
(2S,2'S)-2,2'-(pentane-1,5-diylbis[oxy[(11S,11aS)-11-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8,10(5H)-diyl]carbonyloxymethanediylbenzene-4,1-diyloxycarbonylimino])dipentanedioic acid + 2 H2O
3-hydroxybenzyl 4-hydroxybenzyl (11S,11aS,11'S,11a'S)-8,8'-[pentane-1,5-diylbis(oxy)]bis(11-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-10(5H)-carboxylate) + 2 N-carboxy-L-glutamate
show the reaction diagram
-
-
-
-
?
(4-[bis(2-bromoethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid + H2O
4-[bis(2-bromoethyl)amino]-3,5-difluorobenzoic acid + L-glutamate
show the reaction diagram
(4-[bis(2-iodoethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid + H2O
4-[bis(2-iodoethyl)amino]-3,5-difluorobenzoic acid + L-glutamate
show the reaction diagram
(6R,S)-leucovorin + H2O
N-(6R,S)-5-formyl-5,6,7,8-tetrahydropteroic acid + L-glutamate
show the reaction diagram
-
inactivation of the antitumor drug
-
-
?
4-([bis(2-iodoethyl)amino]-phenyl)oxycarbonyl-L-glutamic acid + H2O
4-[bis(2-iodoethyl)amino]-phenylcarbonate + L-glutamate
show the reaction diagram
-
substrate can be used as a prodrug that is hydrolyzed to yield the active drug in an antibody directed enzyme prodrug therapy
-
-
?
7-hydroxy-methotrexate + H2O
7-hydroxy-4-amino-4-deoxy-N10-methylpteroic acid + L-glutamate
show the reaction diagram
-
-
-
-
?
diethylstilbestrol-glutamate + H2O
diethylstilbestrol + L-glutamate
show the reaction diagram
-
the water soluble substrate can be used as a prodrug that is hydrolyzed to yield the active drug in an antibody directed enzyme prodrug therapy
-
-
?
methotrexate + H2O
4-amino-4-deoxy-N10-methylpteroic acid + L-glutamate
show the reaction diagram
-
highly reduced clearance of the drug in enzyme-defective patients leads to severe neutropenia, mucositis, nephrotoxicity, neurotoxicity, and elevation of liver enzymes in plasma
-
-
?
methotrexate + H2O
4-[(2,4-diamino-6-(pteridinyl)methyl)-methylamino]-benzoic acid + L-glutamate
show the reaction diagram
N-([4-[([[(11S,11aS)-11-hydroxy-7,8-dimethoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-10(5H)-yl]carbonyl]oxy)methyl]phenoxy]carbonyl)-L-glutamic acid + H2O
(11aS)-7,8-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepin-5-one + ?
show the reaction diagram
-
-
-
-
?
N-([4-[([[(11S,11aS)-11-hydroxy-7,8-dimethoxy-5-oxo-2,3,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-10(5H)-yl]carbonyl]oxy)methyl]phenyl]carbamoyl)-L-glutamic acid + H2O
? + L-glutamate
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
weak activator in absence of EDTA
additional information
Mg2+ inhibits the activity of the recombinant enzyme; Mn2+ inhibits the activity of the recombinant enzyme
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(4-[bis(2-chloroethyl)amino]-2,3,5,6-tetrafluorobenzoyl)-L-glutamic acid
-
prodrug, competitive inhibition
(4-[bis(2-iodoethyl)amino]-2,3,5,6-tetrafluorobenzoyl)-L-glutamic acid
-
prodrug, competitive inhibition
2-mercaptoethanol
-
-
4-Aminopteroylaspartic acid
-
hydrolysis of 4-amino-N10-methylpteroylglutamic acid
Ac-Glu
-
inhibition of methotrexate hydrolysis
Ala-Glu
-
inhibition of methotrexate hydrolysis
Benzoyl-Glu
-
inhibition of methotrexate hydrolysis
Benzyloxycarbonyl-Glu
-
inhibition of methotrexate hydrolysis
Benzyloxycarbonyl-Gly-Glu
-
inhibition of methotrexate hydrolysis
Glu-Glu
-
inhibition of methotrexate hydrolysis
Gly-Glu
-
inhibition of methotrexate hydrolysis
His-Glu
-
inhibition of methotrexate hydrolysis
Ile-Glu
-
inhibition of methotrexate hydrolysis
L-aspartic acid
-
weak inhibition of hydrolysis of 4-amino-N10-methylpteroylglutamic acid
L-Glutamic acid
N-Benzyloxycarbonyl glutamic acid
-
hydrolysis of 4-amino-N10-methylpteroylglutamic acid
N-Benzyloxycarbonylaspartic acid
-
hydrolysis of 4-amino-N10-methylpteroylglutamic acid
Phe-Glu
-
inhibition of methotrexate hydrolysis
potassium ascorbate
-
1 mM: harmless, 3 mM: strong inhibition
Pteroic acid
-
-
Tyr-Glu
-
inhibition of methotrexate hydrolysis
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0034
(2-chloroethyl)-(2-(methylsulfoyloxyethyl)amino)aminobenzyl-L-glutamic acid
-
pH 7.3, 37 C
0.0011
(4-[bis(2-bromoethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid
-
pH 7.3, 37 C
0.0012
(4-[bis(2-bromoethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid
-
pH 7.3, 37 C
0.0026
(4-[bis(2-chloroethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid
-
pH 7.3, 37 C
0.0017
(4-[bis(2-chloroethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid
-
pH 7.3, 37 C
0.0028
(4-[bis(2-iodoethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid
-
pH 7.3, 37 C
0.0017
(4-[bis(2-iodoethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid
-
pH 7.3, 37 C
0.58
4-amino-N10-methylpteroylaspartate
-
-
0.104
4-aminopteroylaspartate
-
-
0.12
5-formyltetrahydrofolate
-
-
0.0181
5-formyltetrahydrofolic acid
-
-
0.034
5-methyltetrahydrofolate
-
-
0.0129
5-methyltetrahydrofolic acid
-
-
0.0083
aminopterin
-
-
0.0013
di-tert-butyl 4-[[2'-[N,N-bis(2''-chloroethyl)amino]phenyl]carbamoyloxymethyl]phenyloxycarbonyl-L-glutamic acid
-
-
0.00605
diallyl 4-[N-[4'-bis(2''-chloroethyl)aminophenyl]-N-methylcarbamoyloxymethyl]phenylcarbamoyl-L-glutamic acid
-
-
0.00055
diallyl 4-[[2'-[N,N-bis(2''-chloroethyl)amino]phenyl]carbamoyloxymethyl]phenylcarbamoyl-L-glutamic acid
-
-
0.004
folate
-
-
0.0039 - 0.008
methotrexate
0.0011
pteroylglutamic acid
-
-
0.00171
[4-[bis(2-chloroethyl)amino]-3,5-difluorobenzoyl]-L-glutamic acid
-
-
0.0035
[4-[bis(2-hydroxyethyl)amino]-3,5-difluorobenzoyl]-L-glutamic acid
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
583
(2-chloroethyl)-(2-(methylsulfoyloxyethyl)amino)aminobenzyl-L-glutamic acid
Pseudomonas sp.
-
pH 7.3, 37 C
825
(4-[bis(2-bromoethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid
Pseudomonas sp.
-
pH 7.3, 37 C
827
(4-[bis(2-bromoethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid
Pseudomonas sp.
-
pH 7.3, 37 C
1150
(4-[bis(2-chloroethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid
Pseudomonas sp.
-
pH 7.3, 37 C
732
(4-[bis(2-chloroethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid
Pseudomonas sp.
-
pH 7.3, 37 C
220
(4-[bis(2-iodoethyl)amino]-2,3,5-trifluorobenzoyl)-L-glutamic acid
Pseudomonas sp.
-
pH 7.3, 37 C
1150
(4-[bis(2-iodoethyl)amino]-3,5-difluorobenzoyl)-L-glutamic acid
Pseudomonas sp.
-
pH 7.3, 37 C
5.5
diallyl 4-[N-[4'-bis(2''-chloroethyl)aminophenyl]-N-methylcarbamoyloxymethyl]phenylcarbamoyl-L-glutamic acid
Pseudomonas sp.
-
-
2.9
diallyl 4-[[2'-[N,N-bis(2''-chloroethyl)amino]phenyl]carbamoyloxymethyl]phenylcarbamoyl-L-glutamic acid
Pseudomonas sp.
-
-
732
[4-[bis(2-chloroethyl)amino]-3,5-difluorobenzoyl]-L-glutamic acid
Homo sapiens
-
-
747
[4-[bis(2-hydroxyethyl)amino]-3,5-difluorobenzoyl]-L-glutamic acid
Homo sapiens
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
-
hydrolysis of 5-methyltetrahydrofolate, phosphate buffer
6.5 - 6.8
-
hydrolysis of leucoverin, phosphate buffer
7 - 8
-
hydrolysis of 5-methyltetrahydrofolate, Tris buffer
7.2
-
hydrolysis of methotrexate, folic acid
7.5 - 8
-
hydrolysis of leucoverin, Tris buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.3
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
with methotrexate and 5-methyltetrahydrofolate as substrates, the reaction proceeds equally well between 30C and 42C
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
target cell for gene therapy
Manually annotated by BRENDA team
strong expression in the spikelet meristem
Manually annotated by BRENDA team
strong expression in the epidermal layer of the scutellum of 10-DAP embryos
Manually annotated by BRENDA team
-
target cell for gene therapy
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
41000
SDS-PAGE
41800
-
2 * 41800, SDS-PAGE
42000
-
monomer, determined by immunoblotting
46000
-
2 * 46000, SDS-PAGE
53000
-
gel filtration
92000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop method, crystal structure at 2.5 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
half-life: 4 h
25
-
half-life: 90 min
37
-
half-life: 45 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
activity can increase up to 30% if the enzyme at all stages of purification is stored at -20C
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bovine albumin stabilizes against heat inactivation
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-196C, stable for several months
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-20C, 0.01 M Tris-chloride buffer, pH 7.3, 0.01 mM ZnCl2, stable for several months
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-20C, stable for 1 year
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4C, 0.01 M Tris-chloride buffer, pH 7.3, 0.01 mM ZnCl2, stable for 2 to 3 weeks
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
as His-tagged protein in a single step by Ni2+ charged column chromatography
large scale; strain RS-16
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strain RS-16
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
for expression in Salmonella typhimurium cells using the vector pTrc99A
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into a binary vector for Agrobacterium tumefaciens-mediated transformation
overexpression of a codon-optimized gene in Escherichia coli, vector pET28a, the enzyme is expressed to about 60% of the total host protein and the purification of the recombinant His-tagged protein could be achieved in a single step by Ni2+ charged column chromatography
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Refolding is initiated by rapid 25fold dilution of the denatured protein into refolding buffer (100 mM Tris-HCl, pH 8.5, containing 0.1 mM EDTA and 0.5 M arginine)
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
medicine
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