Cloned (Comment) | Organism |
---|---|
expressed in Sf9 cells using the baculovirus system | Homo sapiens |
expression in baculoviral system | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrial inner membrane | - |
Homo sapiens | 5743 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
42000 | - |
SDS-PAGE | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P26439 | - |
- |
Purification (Comment) | Organism |
---|---|
purified from Sf9 cells in the presence of the low-CMC detergent, Igepal CO 720, using a DEAE ion exchange column | Homo sapiens |
Renatured (Comment) | Organism |
---|---|
between pH 7.4 and 4.5, isoform 3betaHSD2 retains its primarily-helical character with a decrease in alpha-helical content at lower pH values, whereas the beta-sheet content remains unchanged throughout. Titrating the pH back to 7.4 restores the original conformation within 25 min. Metabolic conversion assays indicate peak 3betaHSD2 activity at pH 4.5 with 2fold more progesterone synthesized at pH 4.5 than at pH 3.5 and 7.4. Increasing the 3betaSD2 concentration from 1 to 40 microg results in a 7fold increase in progesterone at pH 4.5, but no change at pH 7.4. Incubation with guanidinum hydrochloride shows a three-step cooperative unfolding of 3betaHSD2 from pH 7.4 to 4.5, possibly due to the native state unfolding to the intermediate ion core state. With further decreases in pH, increasing concentrations of guanidinum hydrochloride lead to rapid two-step unfolding that may represent complete loss of structure | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | metabolic conversion assays indicate peak 3betaHSD2 activity at pH 4.5 with 2fold more progesterone synthesized at pH 4.5 than at pH 3.5 and 7.4. Increasing the 3betaSD2 concentration from 1 to 40 microg results in a 7fold increase in progesterone at pH 4.5, but no change at pH 7.4 | Homo sapiens | ? | - |
? | |
pregnenolone + NAD+ | - |
Homo sapiens | progesterone + NADH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
3beta-hydroxysteroid dehydrogenase | - |
Homo sapiens |
3betaHSD2 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
peak 3betaHSD2 activity at pH 4.5 with 2fold more progesterone synthesized at pH 4.5 than at pH 3.5 and 7.4 | Homo sapiens |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
metabolic conversion assays indicate peak 3betaHSD2 activity at pH 4.5 with 2fold more progesterone synthesized at pH 4.5 than at pH 3.5 and 7.4. Increasing the 3betaSD2 concentration from 1 to 40 microg results in a 7fold increase in progesterone at pH 4.5, but no change at pH 7.4. At pH 4-5, 3betaHSD2 takes on a molten globule conformation that promotes the dual functionality of the enzyme, i.e. 3 beta-hydroxysteroid dehydrogenase and Delta 5->4-isomerase activity | Homo sapiens |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4.5 | 7.4 | circular dichroism shows that between pH 7.4 and 4.5, 3betaHSD2 retains its primarily alpha-helical character with a decrease in beta-helical content at lower pH values, whereas the beta-sheet content remains unchanged throughout. Titrating the pH back to 7.4 restores the original conformation | Homo sapiens |
4.5 | 7.4 | incubation with guanidinum hydrochloride shows a three-step cooperative unfolding of 3betaHSD2 from pH 7.4 to 4.5. With further decreases in pH, increasing concentrations of GdmHCl lead to rapid two-step unfolding that may represent complete loss of structure. Between pH 4 and 5, the two intermediate states appear stable. Stopped-flow kinetics show slower unfolding at around pH 4, where the protein is in a pseudostable state. At pH 4-5, 3betaHSD2 takes on a molten globule conformation that promotes the dual functionality of the enzyme | Homo sapiens |