Literature summary for 1.1.1.248 extracted from

Higashi, Y.; Kutchan, T.M.; Smith, T.J.
Atomic structure of salutaridine reductase from the opium poppy (Papaver somniferum) (2011), J. Biol. Chem., 286, 6532-6541.

Search for more literature for 1.1.1.248

Cloned(Commentary)

Cloned (Comment)	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 (Comment)	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, 4°C, X-ray diffraction structure determination and analysis at 1.9 A resolution	Papaver somniferum

Crystallization (Comment)

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, 4°C, X-ray diffraction structure determination and analysis at 1.9 A resolution

Papaver somniferum

Protein Variants

Protein Variants	Comment	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	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
salutaridine + NADPH + H+	Papaver somniferum	-	salutaridinol + NADP+	-	r

Organism

Organism	UniProt	Comment	Textmining
Papaver somniferum	Q071N0	gene salR	-

Purification (Commentary)

Purification (Comment)	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	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
salutaridine + NADPH + H+	-	Papaver somniferum	salutaridinol + NADP+	-	r

Synonyms

Synonyms	Comment	Organism
SalR	-	Papaver somniferum
salutaridine reductase	-	Papaver somniferum

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	-	Papaver somniferum
NADPH	binding structure, overview	Papaver somniferum

General Information

General Information	Comment	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