Cloned (Comment) | Organism |
---|---|
sequence comparisons, recombinant FLAG-tagged enzyme expression in human HEK-293 cells, recombinant expression of His8-tagged, seleniummethionine-labeled wild-type and mutant enzymes in Escherichia coli strain C43(DE3) | Methylotuvimicrobium alcaliphilum |
Crystallization (Comment) | Organism |
---|---|
purified wild-type and mutant enzymes in complex with NADPH, hanging drop vapour diffusion method, mixing of protein solution containing 2 mM NADPH with reservoir solution containing 0.1 M Tris-HCl, pH 7.0, 0.2 M NH4Ac, and 30% v/v pentaerythritol ethoxylate, 5 days, 20°C, platinum-derivatives are obtained by soaking native crystals for 12 h in mother liquor plus 10 mg/ml K2Pt(NO2)4, X-ray diffraction structure determination and analysis at 2.74-4.3 A resolution, selenium-based single-wavelength anomalous dispersion, modeling | Methylotuvimicrobium alcaliphilum |
Protein Variants | Comment | Organism |
---|---|---|
D363A | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
I151M | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
K406A | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
L304M | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
N359A | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
R395A | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
W352A | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
Y241F | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
Y360A | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
Y407A | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
Y414A | site-directed mutagenesis, inactive mutant | Methylotuvimicrobium alcaliphilum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | integral membrane enzyme, the enzyme contains ten transmembrane segments, TM1-10 | Methylotuvimicrobium alcaliphilum | 16020 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol + NADPH + H+ | Methylotuvimicrobium alcaliphilum | - |
4,4-dimethyl-5alpha-cholesta-8,24-dien-3beta-ol + NADP+ | - |
? | |
4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol + NADPH + H+ | Methylotuvimicrobium alcaliphilum 20Z | - |
4,4-dimethyl-5alpha-cholesta-8,24-dien-3beta-ol + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methylotuvimicrobium alcaliphilum | G4SW86 | - |
- |
Methylotuvimicrobium alcaliphilum 20Z | G4SW86 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His8-tagged, seleniummethionine-labeled wild-type and mutant enzymes from Escherichia coli strain C43(DE3) by nickel affinity chromatography and gel filtration | Methylotuvimicrobium alcaliphilum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol + NADPH + H+ | - |
Methylotuvimicrobium alcaliphilum | 4,4-dimethyl-5alpha-cholesta-8,24-dien-3beta-ol + NADP+ | - |
? | |
4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol + NADPH + H+ | - |
Methylotuvimicrobium alcaliphilum 20Z | 4,4-dimethyl-5alpha-cholesta-8,24-dien-3beta-ol + NADP+ | - |
? | |
4,4-dimethyl-5alpha-cholesta-8,14-dien-3beta-ol + NADPH + H+ | - |
Methylotuvimicrobium alcaliphilum | 4,4-dimethyl-5alpha-cholesta-8-en-3beta-ol + NADP+ | - |
? | |
4,4-dimethyl-5alpha-cholesta-8,14-dien-3beta-ol + NADPH + H+ | - |
Methylotuvimicrobium alcaliphilum 20Z | 4,4-dimethyl-5alpha-cholesta-8-en-3beta-ol + NADP+ | - |
? | |
5alpha-cholesta-8,14-dien-3beta-ol + NADPH + H+ | - |
Methylotuvimicrobium alcaliphilum | 5alpha-cholesta-8-en-3beta-ol + NADP+ | - |
? | |
5alpha-cholesta-8,14-dien-3beta-ol + NADPH + H+ | - |
Methylotuvimicrobium alcaliphilum 20Z | 5alpha-cholesta-8-en-3beta-ol + NADP+ | - |
? | |
additional information | enzyme MaSR1 can reduce the double bond of a cholesterol biosynthetic intermediate analogously to the human enzyme | Methylotuvimicrobium alcaliphilum | ? | - |
? | |
additional information | enzyme MaSR1 can reduce the double bond of a cholesterol biosynthetic intermediate analogously to the human enzyme | Methylotuvimicrobium alcaliphilum 20Z | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme contains ten transmembrane segments (TM1-10). Its catalytic domain comprises the carboxy-terminal half (containing TM6-10) and envelops two interconnected pockets, one of which faces the cytoplasm and houses NADPH, while the other one is accessible from the lipid bilayer | Methylotuvimicrobium alcaliphilum |
Synonyms | Comment | Organism |
---|---|---|
C14SR | - |
Methylotuvimicrobium alcaliphilum |
integral membrane sterol reductase | - |
Methylotuvimicrobium alcaliphilum |
MaSR1 | - |
Methylotuvimicrobium alcaliphilum |
TM7SF2 | - |
Methylotuvimicrobium alcaliphilum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | the binding pocket for NADPH is localized to the C-terminal domain transmembrane segments TM6-10 | Methylotuvimicrobium alcaliphilum |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the sterol reductase family | Methylotuvimicrobium alcaliphilum |
metabolism | the enzyme is involved in the cholesterol biosynthesis pathway | Methylotuvimicrobium alcaliphilum |
additional information | the enzyme contains ten transmembrane segments (TM1-10). Its catalytic domain comprises the carboxy-terminal half (containing TM6-10) and envelops two interconnected pockets, one of which faces the cytoplasm and houses NADPH, while the other one is accessible from the lipid bilayer. The reducing end of NADPH meets the sterol substrate at the juncture of the two pockets | Methylotuvimicrobium alcaliphilum |