BRENDA - Enzyme Database show

Identification of key residues determining isomerohydrolase activity of human RPE65

Takahashi, Y.; Moiseyev, G.; Ma, J.X.; J. Biol. Chem. 289, 6743-26751 (2014)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.1.1.64
recombinant expression of wild-type and mutant enzymes in HEK-293A-LRAT cells
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.1.1.64
C106Y
site-directed mutagenesis, the mutant exhibits 16% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
C106Y/K297G
site-directed mutagenesis, the mutant exhibits 26% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
C330T
site-directed mutagenesis, the mutant exhibits 27% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
I220M
site-directed mutagenesis, the mutant exhibits 38% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
K297G
site-directed mutagenesis, the mutant exhibits 1.7fold increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
K297G/L510M
site-directed mutagenesis, the mutant exhibits 45% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
K297G/N302I
site-directed mutagenesis, the mutant exhibits 7.7% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
K297G/S533A
site-directed mutagenesis, the mutant exhibits 18% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
L510M
site-directed mutagenesis, the mutant exhibits 29% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
additional information
mutant enzymes' recombinant expression levels, overview
Homo sapiens
3.1.1.64
N170K
site-directed mutagenesis, the mutant exhibits 1.6fold increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N170K/C330T
site-directed mutagenesis, the mutant exhibits 15% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N170K/I220M
site-directed mutagenesis, the mutant exhibits 69% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N170K/K297G
site-directed mutagenesis, the mutant exhibits 67% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N170K/Q497P
site-directed mutagenesis, the mutant exhibits 16% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N302I
site-directed mutagenesis, the mutant exhibits 25% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
Q497P
site-directed mutagenesis, the mutant exhibits 26% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
S533A
site-directed mutagenesis, the mutant exhibits 23% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R
site-directed mutagenesis, the mutant exhibits 22% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K
site-directed mutagenesis, the mutant exhibits 39% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/C330T
site-directed mutagenesis, the mutant exhibits 8.4% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/C330T/Q497P
site-directed mutagenesis, the mutant exhibits 23% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/I220M
site-directed mutagenesis, the mutant exhibits 10% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/I220M/Q497P
site-directed mutagenesis, the mutant exhibits 8.6% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/Q497P
site-directed mutagenesis, the mutant exhibits 11% increased activity compared to the wild-type enzyme
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.1.64
0.00104
-
an all-trans-retinyl ester
pH and temperature not specified in the publication
Homo sapiens
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.1.1.64
Homo sapiens
-
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.1.64
an all-trans-retinyl ester + H2O
the enzyme generates predominantly 11-cis-retinol (72.2%) and a minor amount of 13-cis-retinol (27.8%), from all-trans-retinyl ester
730062
Homo sapiens
11-cis-retinol + a fatty acid
-
-
-
?
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.1.64
recombinant expression of wild-type and mutant enzymes in HEK-293A-LRAT cells
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.1.1.64
C106Y
site-directed mutagenesis, the mutant exhibits 16% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
C106Y/K297G
site-directed mutagenesis, the mutant exhibits 26% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
C330T
site-directed mutagenesis, the mutant exhibits 27% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
I220M
site-directed mutagenesis, the mutant exhibits 38% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
K297G
site-directed mutagenesis, the mutant exhibits 1.7fold increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
K297G/L510M
site-directed mutagenesis, the mutant exhibits 45% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
K297G/N302I
site-directed mutagenesis, the mutant exhibits 7.7% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
K297G/S533A
site-directed mutagenesis, the mutant exhibits 18% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
L510M
site-directed mutagenesis, the mutant exhibits 29% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
additional information
mutant enzymes' recombinant expression levels, overview
Homo sapiens
3.1.1.64
N170K
site-directed mutagenesis, the mutant exhibits 1.6fold increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N170K/C330T
site-directed mutagenesis, the mutant exhibits 15% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N170K/I220M
site-directed mutagenesis, the mutant exhibits 69% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N170K/K297G
site-directed mutagenesis, the mutant exhibits 67% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N170K/Q497P
site-directed mutagenesis, the mutant exhibits 16% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
N302I
site-directed mutagenesis, the mutant exhibits 25% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
Q497P
site-directed mutagenesis, the mutant exhibits 26% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
S533A
site-directed mutagenesis, the mutant exhibits 23% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R
site-directed mutagenesis, the mutant exhibits 22% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K
site-directed mutagenesis, the mutant exhibits 39% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/C330T
site-directed mutagenesis, the mutant exhibits 8.4% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/C330T/Q497P
site-directed mutagenesis, the mutant exhibits 23% reduced activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/I220M
site-directed mutagenesis, the mutant exhibits 10% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/I220M/Q497P
site-directed mutagenesis, the mutant exhibits 8.6% increased activity compared to the wild-type enzyme
Homo sapiens
3.1.1.64
T39R/N170K/Q497P
site-directed mutagenesis, the mutant exhibits 11% increased activity compared to the wild-type enzyme
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.1.64
0.00104
-
an all-trans-retinyl ester
pH and temperature not specified in the publication
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.1.64
an all-trans-retinyl ester + H2O
the enzyme generates predominantly 11-cis-retinol (72.2%) and a minor amount of 13-cis-retinol (27.8%), from all-trans-retinyl ester
730062
Homo sapiens
11-cis-retinol + a fatty acid
-
-
-
?
General Information
EC Number
General Information
Commentary
Organism
3.1.1.64
metabolism
the enzyme catalyzes the key reaction in the retinoid visual cycle
Homo sapiens
General Information (protein specific)
EC Number
General Information
Commentary
Organism
3.1.1.64
metabolism
the enzyme catalyzes the key reaction in the retinoid visual cycle
Homo sapiens