Literature summary for 1.1.1.145 extracted from

Prasad, M.; Thomas, J.L.; Whittal, R.M.; Bose, H.S.
Mitochondrial 3beta-hydroxysteroid dehydrogenase enzyme activity requires reversible pH-dependent conformational change at the intermembrane space (2012), J. Biol. Chem., 287, 9534-9546.

Search for more literature for 1.1.1.145

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Sf9 cells using the baculovirus system	Homo sapiens
expression in baculoviral system	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
mitochondrial inner membrane	-	Homo sapiens	5743	-

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
42000	-	SDS-PAGE	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P26439	-	-

Purification (Commentary)

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 (Commentary)

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

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 and Products (Substrate)

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

Synonyms	Comment	Organism
3beta-hydroxysteroid dehydrogenase	-	Homo sapiens
3betaHSD2	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens

pH Optimum

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 Range

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	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