Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary 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.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.248 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)

EC Number Crystallization (Comment) Organism
1.1.1.248 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

EC Number Protein Variants Comment Organism
1.1.1.248 D107A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 F104A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 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
1.1.1.248 I275A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 I275V the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 K186V the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 L185A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 L185S the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 L185V the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 L266A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 M271A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 N272A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 S181A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 T182A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum
1.1.1.248 V106A the mutant shows altered kinetics compared to the wild-type enzyme Papaver somniferum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.248 salutaridine + NADPH + H+ Papaver somniferum
-
 salutaridinol + NADP+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.248 Papaver somniferum Q071N0 gene salR
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.248 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)

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

Synonyms

EC Number Synonyms Comment Organism
1.1.1.248 SalR
-
 Papaver somniferum
1.1.1.248 salutaridine reductase
-
 Papaver somniferum

Cofactor

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

General Information

EC Number General Information Comment Organism
1.1.1.248 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
1.1.1.248 metabolism in the biosynthetic pathway for morphine and codeine, salutaridine is reduced to salutaridinol by salutaridine reductase using NADPH as coenzyme Papaver somniferum
1.1.1.248 additional information modeling of substrate binding and conformation of bound salutaridine, interactions between SalR and its substrate and coenzyme, overview Papaver somniferum