BRENDA - Enzyme Database show

Structural and functional characterization of human microsomal prostaglandin E synthase-1 by computational modeling and site-directed mutagenesis

Huang, X.; Yan, W.; Gao, D.; Tong, M.; Tai, H.H.; Zhan, C.G.; Bioorg. Med. Chem. 14, 3553-3562 (2006)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
5.3.99.3
from human placenta cDNA library, expression in Escherichia coli
Homo sapiens
Crystallization (Commentary)
EC Number
Crystallization
Organism
5.3.99.3
three-dimensional model of substrate-domain and its binding with subsrtates prostaglandin H2 and glutathione. Residue Y130 plays a key role in binding with prostaglandin H2 and in the catalytic process. R110 and T114 interact intensively with the carbonyl tail of prostaglandin H2, whereas Q36 and Q134 only enhance the prostaglandin H2 binding affinity. Prostaglandin H2 interacts with glutathione through hydrogen binding between the peroxy group of prostaglandin H2 and the -SH group of glutathione
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
5.3.99.3
Q134E
fivefold increase in Km value
Homo sapiens
5.3.99.3
Q36E
tenfold increase in Km value
Homo sapiens
5.3.99.3
R110T
18% of wild-type activity
Homo sapiens
5.3.99.3
T114V
21% of wild-type activity
Homo sapiens
5.3.99.3
Y130I
loss of most of the enzymic activity
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.3.99.3
0.13
-
(5Z,13E)-(15S)-9a,11a-epidioxy-15-hydroxyprosta-5,13-dienoate
wild-type, 0C, pH 7.2
Homo sapiens
5.3.99.3
0.734
-
(5Z,13E)-(15S)-9a,11a-epidioxy-15-hydroxyprosta-5,13-dienoate
mutant Q134E, 0C, pH 7.2
Homo sapiens
5.3.99.3
1.61
-
(5Z,13E)-(15S)-9a,11a-epidioxy-15-hydroxyprosta-5,13-dienoate
mutant Q36E, 0C, pH 7.2
Homo sapiens
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
5.3.99.3
membrane
-
Homo sapiens
16020
-
5.3.99.3
microsome
-
Homo sapiens
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
5.3.99.3
Homo sapiens
-
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5.3.99.3
(5Z,13E)-(15S)-9a,11a-epidioxy-15-hydroxyprosta-5,13-dienoate + GSH
-
678584
Homo sapiens
(5Z,13E)-(15S)-11a,15-dihydroxyprosta-5,13-dienoate + GSSG
-
-
-
?
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
5.3.99.3
from human placenta cDNA library, expression in Escherichia coli
Homo sapiens
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
5.3.99.3
three-dimensional model of substrate-domain and its binding with subsrtates prostaglandin H2 and glutathione. Residue Y130 plays a key role in binding with prostaglandin H2 and in the catalytic process. R110 and T114 interact intensively with the carbonyl tail of prostaglandin H2, whereas Q36 and Q134 only enhance the prostaglandin H2 binding affinity. Prostaglandin H2 interacts with glutathione through hydrogen binding between the peroxy group of prostaglandin H2 and the -SH group of glutathione
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
5.3.99.3
Q134E
fivefold increase in Km value
Homo sapiens
5.3.99.3
Q36E
tenfold increase in Km value
Homo sapiens
5.3.99.3
R110T
18% of wild-type activity
Homo sapiens
5.3.99.3
T114V
21% of wild-type activity
Homo sapiens
5.3.99.3
Y130I
loss of most of the enzymic activity
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.3.99.3
0.13
-
(5Z,13E)-(15S)-9a,11a-epidioxy-15-hydroxyprosta-5,13-dienoate
wild-type, 0C, pH 7.2
Homo sapiens
5.3.99.3
0.734
-
(5Z,13E)-(15S)-9a,11a-epidioxy-15-hydroxyprosta-5,13-dienoate
mutant Q134E, 0C, pH 7.2
Homo sapiens
5.3.99.3
1.61
-
(5Z,13E)-(15S)-9a,11a-epidioxy-15-hydroxyprosta-5,13-dienoate
mutant Q36E, 0C, pH 7.2
Homo sapiens
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
5.3.99.3
membrane
-
Homo sapiens
16020
-
5.3.99.3
microsome
-
Homo sapiens
-
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5.3.99.3
(5Z,13E)-(15S)-9a,11a-epidioxy-15-hydroxyprosta-5,13-dienoate + GSH
-
678584
Homo sapiens
(5Z,13E)-(15S)-11a,15-dihydroxyprosta-5,13-dienoate + GSSG
-
-
-
?