Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol
The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogen...
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| Autor*in: |
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| Format: |
E-Artikel |
|---|---|
| Sprache: |
Englisch |
| Erschienen: |
1990 |
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| Reproduktion: |
Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 |
|---|---|
| Übergeordnetes Werk: |
in: The Journal of Steroid Biochemistry and Molecular Biology - Amsterdam : Elsevier, 37(1990), 2, Seite 215-222 |
| Übergeordnetes Werk: |
volume:37 ; year:1990 ; number:2 ; pages:215-222 |
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| 245 | 1 | 0 | |a Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol |
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| 520 | |a The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. | ||
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(DE-627)NLEJ187415145 (DE-599)GBVNLZ187415145 DE-627 ger DE-627 rakwb eng Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol 1990 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Campbell, J.S. oth Karavolas, H.J. oth in The Journal of Steroid Biochemistry and Molecular Biology Amsterdam : Elsevier 37(1990), 2, Seite 215-222 (DE-627)NLEJ18741422X (DE-600)1482780-3 0960-0760 nnns volume:37 year:1990 number:2 pages:215-222 http://linkinghub.elsevier.com/retrieve/pii/0960-0760(90)90329-J GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 37 1990 2 215-222 |
| spelling |
(DE-627)NLEJ187415145 (DE-599)GBVNLZ187415145 DE-627 ger DE-627 rakwb eng Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol 1990 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Campbell, J.S. oth Karavolas, H.J. oth in The Journal of Steroid Biochemistry and Molecular Biology Amsterdam : Elsevier 37(1990), 2, Seite 215-222 (DE-627)NLEJ18741422X (DE-600)1482780-3 0960-0760 nnns volume:37 year:1990 number:2 pages:215-222 http://linkinghub.elsevier.com/retrieve/pii/0960-0760(90)90329-J GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 37 1990 2 215-222 |
| allfields_unstemmed |
(DE-627)NLEJ187415145 (DE-599)GBVNLZ187415145 DE-627 ger DE-627 rakwb eng Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol 1990 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Campbell, J.S. oth Karavolas, H.J. oth in The Journal of Steroid Biochemistry and Molecular Biology Amsterdam : Elsevier 37(1990), 2, Seite 215-222 (DE-627)NLEJ18741422X (DE-600)1482780-3 0960-0760 nnns volume:37 year:1990 number:2 pages:215-222 http://linkinghub.elsevier.com/retrieve/pii/0960-0760(90)90329-J GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 37 1990 2 215-222 |
| allfieldsGer |
(DE-627)NLEJ187415145 (DE-599)GBVNLZ187415145 DE-627 ger DE-627 rakwb eng Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol 1990 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Campbell, J.S. oth Karavolas, H.J. oth in The Journal of Steroid Biochemistry and Molecular Biology Amsterdam : Elsevier 37(1990), 2, Seite 215-222 (DE-627)NLEJ18741422X (DE-600)1482780-3 0960-0760 nnns volume:37 year:1990 number:2 pages:215-222 http://linkinghub.elsevier.com/retrieve/pii/0960-0760(90)90329-J GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 37 1990 2 215-222 |
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(DE-627)NLEJ187415145 (DE-599)GBVNLZ187415145 DE-627 ger DE-627 rakwb eng Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol 1990 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Campbell, J.S. oth Karavolas, H.J. oth in The Journal of Steroid Biochemistry and Molecular Biology Amsterdam : Elsevier 37(1990), 2, Seite 215-222 (DE-627)NLEJ18741422X (DE-600)1482780-3 0960-0760 nnns volume:37 year:1990 number:2 pages:215-222 http://linkinghub.elsevier.com/retrieve/pii/0960-0760(90)90329-J GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 37 1990 2 215-222 |
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purification of the nadph:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol |
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Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol |
| abstract |
The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. |
| abstractGer |
The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. |
| abstract_unstemmed |
The NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase (3α- HSOR) [EC 1.1.1.50] which catalyzes the reversible conversion of 5α-pregnane-3,20-dione (5α-dihydroprogesterone; 5α-DHP) to 3α-hydroxy-5α-pregnan-20-one (3α-,5α-tetrahydro-progesterone; 3α,5α-THP) was purified to apparent homogeneity from female rat anterior pituitary cytosol by a three step micro-purification. Specific activity of purified 3α-HSOR was enriched 438-fold from that in pituitary cytosol using successive ion exchange, chromatofocusing and affinity column chromatography purification steps. 3α-HSOR appears to be a monomer with an approximate molecular weight of 36 kDa and an isoelectric point of about 5.75. The purified enzyme appears as a single protein staining band (36 kDa) when examined by polyacrylamide gel electrophoresis and with both silver or Coomassie blue staining. Under non-dissociating electrophoretic conditions, all of the 3α-HSOR activity co-migrated with the 36 kDa protein staining band. The purified enzyme in the presence of the preferred cofactor, NADPH, has an apparent K"m for 5α-DHP of 82 nM and a V"m"a"x of 1.2 μmol of 3α, 5α-THP formed per mg protein/30 min. The K"m for NADPH was 0.71 μM. In the oxidative direction, the enzyme in the presence of NADP^+ has a K"m for 3α,5α-THP of 1.4 μM and a V"m"a"x of 9.7 μmol of 5α-DHP formed per mg protein/30 min. The K"m for NADP^+ was 1.6 μM. |
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Purification of the NADPH:5α-dihydroprogesterone 3α-hydroxysteroid oxidoreductase from female rat pituitary cytosol |
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7.401164 |