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 (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 | 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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
salutaridine + NADPH + H+ | Papaver somniferum | - |
salutaridinol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Papaver somniferum | Q071N0 | gene salR | - |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
salutaridine + NADPH + H+ | - |
Papaver somniferum | salutaridinol + NADP+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
SalR | - |
Papaver somniferum |
salutaridine reductase | - |
Papaver somniferum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Papaver somniferum | |
NADPH | binding structure, overview | Papaver somniferum |
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 |