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show all sequences of 1.1.1.248

Atomic structure of salutaridine reductase from the opium poppy (Papaver somniferum)

Higashi, Y.; Kutchan, T.M.; Smith, T.J.; J. Biol. Chem. 286, 6532-6541 (2011)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene salR, expression of N-terminally His-tagged enzyme in Escherichia coli, a selenomethionine-substituted SalR is produced by inhibition of the methionine biosynthetic pathway with the same expression vector and Escherichia coli strain used for expression of wild-type SalR
Papaver somniferum
Crystallization (Commentary)
Crystallization
Organism
purified recombinant detagged wild-type and selenomethionine-substituted SalR, hanging drop vapour diffsion method, mixing of 0.002 ml of 6 mg/ml protein in 20 mM Tris buffer, pH 7.5, containing 150 mM NaCl, 5 mM 2-mercaptoethanol, and 4 mM NADPH, with 0.002 ml of reservoir solution containing 0.1 M MES, pH 6.0-6.6, 1.9 M ammonium sulfate, 5% v/v PEG 400, 0.1 M LiCl, and 3% v/v glycerol, 3 weeks, 4C, X-ray diffraction structure determination and analysis at 1.9 A resolution
Papaver somniferum
Engineering
Amino acid exchange
Commentary
Organism
D107A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
F104A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
F104A/I275A
substitution of Phe104 in the substrate-binding pocket, and Ile275 under the flap domain, the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
I275A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
I275V
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
K186V
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
L185A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
L185S
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
L185V
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
L266A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
M271A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
N272A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
S181A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
T182A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
V106A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
salutaridine + NADPH + H+
Papaver somniferum
-
salutaridinol + NADP+
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Papaver somniferum
Q071N0
gene salR
-
Purification (Commentary)
Commentary
Organism
recombinant N-terminally His-tagged wild-type and selenomethionine-substituted SalR from Escherichia coli by cobalt affinity chromatography, cleavage of the N-terminal His-tag by thrombin, and gel filtration
Papaver somniferum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
salutaridine + NADPH + H+
-
725460
Papaver somniferum
salutaridinol + NADP+
-
-
-
r
Cofactor
Cofactor
Commentary
Organism
Structure
NADP+
-
Papaver somniferum
NADPH
binding structure, overview
Papaver somniferum
Cloned(Commentary) (protein specific)
Commentary
Organism
gene salR, expression of N-terminally His-tagged enzyme in Escherichia coli, a selenomethionine-substituted SalR is produced by inhibition of the methionine biosynthetic pathway with the same expression vector and Escherichia coli strain used for expression of wild-type SalR
Papaver somniferum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADP+
-
Papaver somniferum
NADPH
binding structure, overview
Papaver somniferum
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified recombinant detagged wild-type and selenomethionine-substituted SalR, hanging drop vapour diffsion method, mixing of 0.002 ml of 6 mg/ml protein in 20 mM Tris buffer, pH 7.5, containing 150 mM NaCl, 5 mM 2-mercaptoethanol, and 4 mM NADPH, with 0.002 ml of reservoir solution containing 0.1 M MES, pH 6.0-6.6, 1.9 M ammonium sulfate, 5% v/v PEG 400, 0.1 M LiCl, and 3% v/v glycerol, 3 weeks, 4C, X-ray diffraction structure determination and analysis at 1.9 A resolution
Papaver somniferum
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D107A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
F104A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
F104A/I275A
substitution of Phe104 in the substrate-binding pocket, and Ile275 under the flap domain, the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
I275A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
I275V
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
K186V
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
L185A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
L185S
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
L185V
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
L266A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
M271A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
N272A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
S181A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
T182A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
V106A
the mutant shows altered kinetics compared to the wild-type enzyme
Papaver somniferum
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
salutaridine + NADPH + H+
Papaver somniferum
-
salutaridinol + NADP+
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant N-terminally His-tagged wild-type and selenomethionine-substituted SalR from Escherichia coli by cobalt affinity chromatography, cleavage of the N-terminal His-tag by thrombin, and gel filtration
Papaver somniferum
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
salutaridine + NADPH + H+
-
725460
Papaver somniferum
salutaridinol + NADP+
-
-
-
r
General Information
General Information
Commentary
Organism
evolution
the enzyme is a member of the short chain dehydrogenase/reductase family of enzymes. The nicotinamide moiety and the substrate-binding pocket are covered by a loop (residues 265-279), on top of which lies a large flap-like domain (residues 105-140). This configuration appears to be a combination of the two common structural themes found in other members of the short chain dehydrogenase/reductase family.
Papaver somniferum
metabolism
in the biosynthetic pathway for morphine and codeine, salutaridine is reduced to salutaridinol by salutaridine reductase using NADPH as coenzyme
Papaver somniferum
additional information
modeling of substrate binding and conformation of bound salutaridine, interactions between SalR and its substrate and coenzyme, overview
Papaver somniferum
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme is a member of the short chain dehydrogenase/reductase family of enzymes. The nicotinamide moiety and the substrate-binding pocket are covered by a loop (residues 265-279), on top of which lies a large flap-like domain (residues 105-140). This configuration appears to be a combination of the two common structural themes found in other members of the short chain dehydrogenase/reductase family.
Papaver somniferum
metabolism
in the biosynthetic pathway for morphine and codeine, salutaridine is reduced to salutaridinol by salutaridine reductase using NADPH as coenzyme
Papaver somniferum
additional information
modeling of substrate binding and conformation of bound salutaridine, interactions between SalR and its substrate and coenzyme, overview
Papaver somniferum
Other publictions for EC 1.1.1.248
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
720707
Wijekoon
Systematic knockdown of morphi ...
Papaver somniferum
Plant J.
69
1052-1063
2012
-
-
1
-
1
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
725460
Higashi
Atomic structure of salutaridi ...
Papaver somniferum
J. Biol. Chem.
286
6532-6541
2011
-
-
1
1
15
-
-
-
-
-
-
1
-
3
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
1
15
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
3
3
-
-
-
710745
Higashi
Crystallization and preliminar ...
Papaver somniferum
Acta Crystallogr. Sect. F
66
163-166
2010
-
-
1
1
-
-
-
-
-
-
-
-
-
4
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
698937
Ziegler
Removal of substrate inhibitio ...
Papaver somniferum
J. Biol. Chem.
284
26758-26767
2009
-
-
1
-
12
-
2
26
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
13
-
-
-
1
1
-
-
-
-
1
1
-
12
-
-
2
1
26
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
13
-
-
-
-
-
-
-
-
-
-
700676
Ziegler
Evolution of morphine biosynth ...
Papaver arenarium, Papaver bracteatum, Papaver pilosum, Papaver pyrenaicum, Papaver somniferum
Phytochemistry
70
1696-1707
2009
-
-
1
-
-
-
-
-
-
-
-
-
-
8
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
5
-
-
-
-
-
1
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
700683
Kempe
RNAi suppression of the morphi ...
Papaver somniferum
Phytochemistry
70
579-589
2009
-
-
1
-
-
-
1
-
1
-
-
-
-
3
-
-
1
-
-
1
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
689571
Geissler
Molecular modeling and site-di ...
Papaver bracteatum
Plant Physiol.
143
1493-1503
2007
-
-
1
1
13
-
1
6
-
-
2
2
-
4
-
-
1
-
-
-
-
-
4
1
1
-
-
5
2
1
-
4
1
1
-
-
-
1
4
1
13
-
-
1
1
6
-
-
2
2
-
-
-
1
-
-
-
-
4
1
1
-
-
5
2
1
-
1
-
-
-
-
-
-
670562
Ziegler
Comparative transcript and alk ...
Papaver somniferum, Papaver somniferum salR
Plant J.
48
177-192
2006
-
-
1
-
-
-
-
4
-
-
3
4
-
6
-
-
1
-
-
-
-
-
8
1
1
1
-
-
2
2
-
4
-
1
-
-
-
1
4
-
-
-
-
-
-
4
-
-
3
4
-
-
-
1
-
-
-
-
8
1
1
1
-
-
2
2
-
1
-
-
-
-
-
-
9554
Gerardy
-
Purification and characterizat ...
no activity in Papaver alpinum, no activity in Papaver atlanticum, no activity in Papaver dubium, no activity in Papaver feddei, no activity in Papaver lateritium, no activity in Papaver oreophilum, no activity in Papaver orientale, no activity in Papaver persicum, no activity in Papaver pilosum, no activity in Papaver rhoeas, no activity in Papaver rupifragum, no activity in Papaver strigosum, no activity in Papaver tauricula, no activity in Papaver triniifolium, Papaver bracteatum, Papaver somniferum
Phytochemistry
34
125-132
1993
-
-
-
-
-
-
-
4
1
-
1
1
-
16
-
-
1
1
-
6
1
1
2
1
1
-
-
-
2
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
1
-
1
1
-
-
-
1
-
6
1
1
2
1
1
-
-
-
2
2
1
-
-
-
-
-
-
-