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Literature summary extracted from

  • Hartfield, K.A.; Stout, C.D.; Annalora, A.J.
    The novel purification and biochemical characterization of a reversible CYP24A1:adrenodoxin complex (2013), J. Steroid Biochem. Mol. Biol., 136, 47-53.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.16 gene CYP24A1, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain DH5alpha F'IQ Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
1.14.15.16 additional information development of truncated expression constructs for rat DELTA51CYP24A1. Adrenodoxin binding enhances the stability of the enzyme-substrate complex, despite lowering the ligand binding affinity of the free enzyme for calcitriol over 9fold. Truncation of CYP24A1's flexible N-terminus (DELTA51) improves the enzyme's ability to recruit substrate, without altering adrenodoxin's ability to stabilize the ligand-bound form Rattus norvegicus
1.14.15.16 S57D site-directed mutagenesis Rattus norvegicus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.15.16 5-cyclohexylpentyl beta-D-maltoside
-
Rattus norvegicus
1.14.15.16 6-cyclohexylhexyl beta-D-maltoside
-
Rattus norvegicus
1.14.15.16 7-cyclohexylheptyl beta-D-maltoside
-
Rattus norvegicus
1.14.15.16 CHAPS inhibits ligand binding to the CYP24A1:adrenodoxin complex at concentrations well below their reported critical micelle concentration (CMC) values Rattus norvegicus
1.14.15.16 Facade-EM
-
Rattus norvegicus
1.14.15.16 Fos-choline-14
-
Rattus norvegicus
1.14.15.16 additional information detergent inhibition properties of the purified enzyme-adrenodoxin cofactor complex bound to calcitriol, overview Rattus norvegicus
1.14.15.16 n-dodecyl beta-D-maltoside
-
Rattus norvegicus
1.14.15.16 nonyl-beta-D-glucoside
-
Rattus norvegicus
1.14.15.16 octa ethylene glycol monododecyl ether
-
Rattus norvegicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.15.16 55000
-
x * 55000, about, recombinant His6-tagged enzyme, SDS-PAGE Rattus norvegicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.15.16 calcitriol + 2 reduced adrenodoxin + 2 H+ + O2 Rattus norvegicus
-
calcitetrol + 2 oxidized adrenodoxin + H2O
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.14.15.16 Rattus norvegicus
-
gene CYP24A1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.15.16 recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain DH5alpha F'IQ by nickel affinity chromatography and gel filtration, copurification with added recombinant bovine adrenodoxin for enzyme-cofctor complex preparation Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.16 calcitriol + 2 reduced adrenodoxin + 2 H+ + O2
-
Rattus norvegicus calcitetrol + 2 oxidized adrenodoxin + H2O
-
?
1.14.15.16 calcitriol + 2 reduced adrenodoxin + 2 H+ + O2 recombinant truncated DELTA108 adrenodoxin from Bos taurus is used Rattus norvegicus calcitetrol + 2 oxidized adrenodoxin + H2O
-
?
1.14.15.16 additional information detailed analysis of the reversible complex between recombinant truncated enzyme DELT51CYP24A1 and recombinant truncated DELTA108adrenodoxin, kinetics, overview. Adrenodoxin binding enhances the stability of the enzyme-substrate complex, despite lowering the ligand binding affinity of the free enzyme for calcitriol over 9fold Rattus norvegicus ?
-
?

Subunits

EC Number Subunits Comment Organism
1.14.15.16 ? x * 55000, about, recombinant His6-tagged enzyme, SDS-PAGE Rattus norvegicus

Synonyms

EC Number Synonyms Comment Organism
1.14.15.16 CYP24A1
-
Rattus norvegicus

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.15.16 adrenodoxin
-
Rattus norvegicus
1.14.15.16 heme
-
Rattus norvegicus
1.14.15.16 NADPH
-
Rattus norvegicus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.14.15.16 0.29
-
n-dodecyl beta-D-maltoside pH and temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.29
-
octa ethylene glycol monododecyl ether pH and temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.37
-
Facade-EM pH and temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.95
-
7-cyclohexylheptyl beta-D-maltoside pH and temperature not specified in the publication Rattus norvegicus
1.14.15.16 1.05
-
5-cyclohexylpentyl beta-D-maltoside pH and temperature not specified in the publication Rattus norvegicus
1.14.15.16 1.67
-
Fos-choline-14 pH and temperature not specified in the publication Rattus norvegicus
1.14.15.16 1.88
-
6-cyclohexylhexyl beta-D-maltoside pH and temperature not specified in the publication Rattus norvegicus
1.14.15.16 3.65
-
CHAPS pH and temperature not specified in the publication Rattus norvegicus
1.14.15.16 6.9
-
n-dodecyl beta-D-maltoside pH and temperature not specified in the publication Rattus norvegicus