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Literature summary for 1.14.15.16 extracted from
- The novel purification and biochemical characterization of a reversible CYP24A1:adrenodoxin complex (2013), J. Steroid Biochem. Mol. Biol., 136, 47-53.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| 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
| Protein Variants | Comment | Organism |
|---|---|---|
| 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 |
| S57D | site-directed mutagenesis | Rattus norvegicus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 5-cyclohexylpentyl beta-D-maltoside | - |
Rattus norvegicus |  |
| 6-cyclohexylhexyl beta-D-maltoside | - |
Rattus norvegicus | |
| 7-cyclohexylheptyl beta-D-maltoside | - |
Rattus norvegicus | |
| CHAPS | inhibits ligand binding to the CYP24A1:adrenodoxin complex at concentrations well below their reported critical micelle concentration (CMC) values | Rattus norvegicus | |
| Facade-EM | - |
Rattus norvegicus | |
| Fos-choline-14 | - |
Rattus norvegicus | |
| additional information | detergent inhibition properties of the purified enzyme-adrenodoxin cofactor complex bound to calcitriol, overview | Rattus norvegicus | |
| n-dodecyl beta-D-maltoside | - |
Rattus norvegicus | |
| nonyl-beta-D-glucoside | - |
Rattus norvegicus | |
| octa ethylene glycol monododecyl ether | - |
Rattus norvegicus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 55000 | - |
x * 55000, about, recombinant His6-tagged enzyme, SDS-PAGE | Rattus norvegicus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| calcitriol + 2 reduced adrenodoxin + 2 H+ + O2 | Rattus norvegicus | - |
calcitetrol + 2 oxidized adrenodoxin + H2O | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Rattus norvegicus | - |
gene CYP24A1 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| 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)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| calcitriol + 2 reduced adrenodoxin + 2 H+ + O2 | - |
Rattus norvegicus | calcitetrol + 2 oxidized adrenodoxin + H2O | - |
? | |
| calcitriol + 2 reduced adrenodoxin + 2 H+ + O2 | recombinant truncated DELTA108 adrenodoxin from Bos taurus is used | Rattus norvegicus | calcitetrol + 2 oxidized adrenodoxin + H2O | - |
? | |
| 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
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 55000, about, recombinant His6-tagged enzyme, SDS-PAGE | Rattus norvegicus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CYP24A1 | - |
Rattus norvegicus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| adrenodoxin | - |
Rattus norvegicus | |
| heme | - |
Rattus norvegicus | |
| NADPH | - |
Rattus norvegicus | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.29 | - |
n-dodecyl beta-D-maltoside | pH and temperature not specified in the publication | Rattus norvegicus | |
| 0.29 | - |
octa ethylene glycol monododecyl ether | pH and temperature not specified in the publication | Rattus norvegicus | |
| 0.37 | - |
Facade-EM | pH and temperature not specified in the publication | Rattus norvegicus | |
| 0.95 | - |
7-cyclohexylheptyl beta-D-maltoside | pH and temperature not specified in the publication | Rattus norvegicus | |
| 1.05 | - |
5-cyclohexylpentyl beta-D-maltoside | pH and temperature not specified in the publication | Rattus norvegicus | |
| 1.67 | - |
Fos-choline-14 | pH and temperature not specified in the publication | Rattus norvegicus | |
| 1.88 | - |
6-cyclohexylhexyl beta-D-maltoside | pH and temperature not specified in the publication | Rattus norvegicus | |
| 3.65 | - |
CHAPS | pH and temperature not specified in the publication | Rattus norvegicus | |
| 6.9 | - |
n-dodecyl beta-D-maltoside | pH and temperature not specified in the publication | Rattus norvegicus | |
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