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

BRENDA support

Literature summary for 1.3.1.70 extracted from

  • Li, X.; Roberti, R.; Blobel, G.
    Structure of an integral membrane sterol reductase from Methylomicrobium alcaliphilum (2015), Nature, 517, 104-107 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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 (Commentary)

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

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

Localization Comment Organism GeneOntology No. Textmining
membrane integral membrane enzyme, the enzyme contains ten transmembrane segments, TM1-10 Methylotuvimicrobium alcaliphilum 16020
-

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Methylotuvimicrobium alcaliphilum G4SW86
-
-
Methylotuvimicrobium alcaliphilum 20Z G4SW86
-
-

Purification (Commentary)

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 and Products (Substrate)

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

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

Synonyms Comment Organism
C14SR
-
Methylotuvimicrobium alcaliphilum
integral membrane sterol reductase
-
Methylotuvimicrobium alcaliphilum
MaSR1
-
Methylotuvimicrobium alcaliphilum
TM7SF2
-
Methylotuvimicrobium alcaliphilum

Cofactor

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

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