BRENDA - Enzyme Database show

Purified RPE65 shows isomerohydrolase activity after reassociation with a phospholipid membrane

Nikolaeva, O.; Takahashi, Y.; Moiseyev, G.; Ma, J.X.; FEBS J. 276, 3020-3030 (2009)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.1.1.64
expressed in 293A-LRAT cells
Gallus gallus
General Stability
EC Number
General Stability
Organism
3.1.1.64
0.1% (w/v) CHAPS is optimal for solubilizing RPE65 while preserving its enzymatic activity
Gallus gallus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.1.64
CHAPS
the addition of 0.5% (w/v) CHAPS to the assay system almost completely abolishes the 11-cis-retinol formation
Gallus gallus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.1.64
0.0037
-
all-trans-retinyl palmitate
purified enzyme, in 10 mM BTP buffer, pH 8.0, 100 mM NaCl, at 37°C
Gallus gallus
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.1.64
membrane
-
Gallus gallus
16020
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.1.1.64
Gallus gallus
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.1.1.64
Ni-NTA column chromatography
Gallus gallus
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
3.1.1.64
retinal pigment epithelium
-
Gallus gallus
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.1.64
all-trans-retinyl ester + H2O
all-trans-retinyl ester must be incorporated into the phospholipid membrane to serve as a substrate for isomerohydrolase RPE65
714951
Gallus gallus
11-cis-retinol + a fatty acid
-
-
-
?
3.1.1.64
all-trans-retinyl palmitate + H2O
high affinity substrate
714951
Gallus gallus
11-cis-retinol + palmitate
-
-
-
?
3.1.1.64
additional information
neither retinyl ester nor 11-cis-retinol is produced after incubation of all-trans-retinol with the purified RPE65 (not an intrinsic substrate for RPE65)
714951
Gallus gallus
?
-
-
-
-
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.1.64
0.000145
-
all-trans-retinyl palmitate
purified enzyme, in 10 mM BTP buffer, pH 8.0, 100 mM NaCl, at 37°C
Gallus gallus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.1.64
expressed in 293A-LRAT cells
Gallus gallus
General Stability (protein specific)
EC Number
General Stability
Organism
3.1.1.64
0.1% (w/v) CHAPS is optimal for solubilizing RPE65 while preserving its enzymatic activity
Gallus gallus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.1.64
CHAPS
the addition of 0.5% (w/v) CHAPS to the assay system almost completely abolishes the 11-cis-retinol formation
Gallus gallus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.1.64
0.0037
-
all-trans-retinyl palmitate
purified enzyme, in 10 mM BTP buffer, pH 8.0, 100 mM NaCl, at 37°C
Gallus gallus
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.1.64
membrane
-
Gallus gallus
16020
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.1.64
Ni-NTA column chromatography
Gallus gallus
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
3.1.1.64
retinal pigment epithelium
-
Gallus gallus
-
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
all-trans-retinyl ester + H2O
all-trans-retinyl ester must be incorporated into the phospholipid membrane to serve as a substrate for isomerohydrolase RPE65
714951
Gallus gallus
11-cis-retinol + a fatty acid
-
-
-
?
3.1.1.64
all-trans-retinyl palmitate + H2O
high affinity substrate
714951
Gallus gallus
11-cis-retinol + palmitate
-
-
-
?
3.1.1.64
additional information
neither retinyl ester nor 11-cis-retinol is produced after incubation of all-trans-retinol with the purified RPE65 (not an intrinsic substrate for RPE65)
714951
Gallus gallus
?
-
-
-
-
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.1.64
0.000145
-
all-trans-retinyl palmitate
purified enzyme, in 10 mM BTP buffer, pH 8.0, 100 mM NaCl, at 37°C
Gallus gallus
General Information
EC Number
General Information
Commentary
Organism
3.1.1.64
physiological function
RPE65 is the isomerohydrolase of the visual cycle
Gallus gallus
General Information (protein specific)
EC Number
General Information
Commentary
Organism
3.1.1.64
physiological function
RPE65 is the isomerohydrolase of the visual cycle
Gallus gallus