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Literature summary extracted from
- The engineered, cytosolic form of human type I 3beta-hydroxysteroid dehydrogenase/isomerase: purification, characterization and crystallization (2001), J. Mol. Endocrinol., 27, 77-83.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.1.1.145 | overexpression with his6-tag, using a baculovirus expression system | Homo sapiens |
| 5.3.3.1 | insect Sf9 cells transfected with recombinant baculovirus | Homo sapiens |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 1.1.1.145 | modified enzyme | Homo sapiens |
| 5.3.3.1 | hanging-drop vapor diffusion method. The inability to crystallize the detergent-solubilized, wild-type microsomal enzyme is overcome by engineering a cytosolic form of this protein. Modified enzyme in which the membrane-spanning domain, residues 283-310 of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells | Homo sapiens |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.3.3.1 | DELTA283-310 | the molecular weight of the subunit is 38800 Da compared to 42000 Da for the wild-type enzyme | Homo sapiens |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.1.1.145 | 0.0034 | - |
NAD+ | wild type enzyme | Homo sapiens |  |
| 1.1.1.145 | 0.0035 | - |
NAD+ | modified enzyme | Homo sapiens | |
| 1.1.1.145 | 0.0037 | - |
dehydroepiandrosterone | wild type enzyme | Homo sapiens | |
| 1.1.1.145 | 0.0045 | - |
dehydroepiandrosterone | modified enzyme | Homo sapiens | |
| 5.3.3.1 | 0.0252 | - |
5-androstene-3,17-dione | 22°C, pH 7.4, truncated cytosolic enzyme DEALTA283-310 | Homo sapiens | |
| 5.3.3.1 | 0.0279 | - |
5-androstene-3,17-dione | 22°C, pH 7.4, wild-type enzyme | Homo sapiens | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 5.3.3.1 | cytosol | modified enzyme in which the membrane-spanning domain, residues 283-310, of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells | Homo sapiens | 5829 | - |
| 5.3.3.1 | membrane | integral membrane protein | Homo sapiens | 16020 | - |
| 5.3.3.1 | microsome | - |
Homo sapiens | - |
- |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.1.1.145 | 42000 | - |
42000, SDS-PAGE | Homo sapiens |
| 5.3.3.1 | 38800 | - |
x * 38800, truncated soluble enzyme DEALTA283-310 | Homo sapiens |
| 5.3.3.1 | 42000 | - |
x * 42000, wild-type enzyme | Homo sapiens |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.1.1.145 | pregnenolone + NAD(P)+ | Homo sapiens | - |
pregn-5-en-3,20-dione + NAD(P)H + H+ | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.1.1.145 | Homo sapiens | - |
3beta-HSD I | - |
| 5.3.3.1 | Homo sapiens | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.3.3.1 | modified enzyme in which the membrane-spanning domain, residues 283-310, of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells | Homo sapiens |
Source Tissue
| EC Number | Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|---|
| 1.1.1.145 | placenta | 3beta-HSD I | Homo sapiens | - |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.1.1.145 | 0.0524 | - |
- |
Homo sapiens |
| 5.3.3.1 | 0.622 | - |
reaction with 5-androsten-3,17-dione, truncated cytosolic enzyme DELTA283-310 | Homo sapiens |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.1.1.145 | 3beta,17alpha-dihydroxy-5-pregnen-20-one + NAD(P)+ | - |
Homo sapiens | 17alpha-hydroxypregn-5-ene-3,20-dione + NAD(P)H | - |
? | |
| 1.1.1.145 | pregnenolone + NAD(P)+ | - |
Homo sapiens | pregn-5-en-3,20-dione + NAD(P)H + H+ | - |
? | |
| 5.3.3.1 | 5-Androstene-3,17-dione | - |
Homo sapiens | 4-Androstene-3,17-dione | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.1.1.145 | ? | 42000, SDS-PAGE | Homo sapiens |
| 5.3.3.1 | ? | x * 38800, truncated soluble enzyme DEALTA283-310 | Homo sapiens |
| 5.3.3.1 | ? | x * 42000, wild-type enzyme | Homo sapiens |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.1.1.145 | NAD+ | - |
Homo sapiens | |
| 5.3.3.1 | NADH | the isomerase activity of the wild-type enzyme and the deletion mutant enzyme DELTA283-310 requires allosteric activation by NADH, Km for wild-type enzyme and deletion mutant enzyme is 0.0046 mM | Homo sapiens | |
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