Purification and properties of testicular 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase
Through the treatment of rat testicular microsomes with sodium cholate, 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase (abbreviated as the 3β -hydroxysteroid dehydrogenase and isomerase, respectively) were solubilized, and then purified by DEAE and hydroxylapatite column chromatogr...
Gespeichert in:
| Veröffentlicht in: | Journal of steroid biochemistry 1986-10, Vol.25 (4), p.555-560 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 560 |
|---|---|
| container_issue | 4 |
| container_start_page | 555 |
| container_title | Journal of steroid biochemistry |
| container_volume | 25 |
| creator | Hiroko, Ishii-Ohba Hiroshi, Inano Bun-Ichi, Tamaoki |
| description | Through the treatment of rat testicular microsomes with sodium cholate, 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase (abbreviated as the 3β -hydroxysteroid dehydrogenase and isomerase, respectively) were solubilized, and then purified by DEAE and hydroxylapatite column chromatographies. The findings were as follows:
1. With this purification procedure, the 3β-hydroxysteroid dehydrogenase activity could not be separated from the isomerase.
2. For 3-oxo-4-ene-steroid formation from 3β-hydroxy-5-ene-steroids, NAD
+ was required as a cofactor. While the 3β-hydroxysteroid dehydrogenase required NAD
+, the isomerase also required NAD
+ or its reduced form, in contrast to the microbial enzyme.
3. On treatment of the purified enzyme with 5'-
p-fluorosulfonyl-benzoyladenosine (FSBA), both enzyme activities were markedly reduced.
4. The enzyme, affinity labeled with [adenine-8-
14C]FSBA, showed a mol. wt of 46.8 K.
5. During 4-androstenedione production from DHA, 5-androstenedione was detected as an intermediate. |
| doi_str_mv | 10.1016/0022-4731(86)90402-4 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_77135317</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>0022473186904024</els_id><sourcerecordid>77135317</sourcerecordid><originalsourceid>FETCH-LOGICAL-c386t-8dc8fdc0e045d8371384d938ae66fff6e68d31f600051e69f0825cedc24e6f753</originalsourceid><addsrcrecordid>eNp9kM1OxCAUhVloxnH0DTTpwhhdVKFQSjcmZuJfYqILXROEi6KdMkJrnNfyQXwm6cxklm4g3HPuCedD6IDgM4IJP8e4KHJWUXIi-GmNGU6vLTTejHfQbozvGJNasGKERkXNyroqxujjsQ_OOq0659tMtSabBz-H0DmImbdZB7Fzum9UyOjvT_62MMF_L_Iyhxby2EHwzmQGlvNXaFWEZchKZ8OZuehnEJKyh7ataiLsr-8Jer6-epre5vcPN3fTy_tcU8G7XBgtrNEYMCuNoBWhgpmaCgWcW2s5cGEosRxjXBLgtcWiKDUYXTDgtirpBB2vclOVzz4VkDMXNTSNasH3UVYpsqSkSka2MurgYwxg5Ty4mQoLSbAcuMoBoBwASsHlkqtkae1wnd-_zMBsltZQk3601lXUqrFBtdrFjU0QUnGBk-1iZYPE4stBkFE7aFMTF0B30nj3_z_-AHP-lyQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>77135317</pqid></control><display><type>article</type><title>Purification and properties of testicular 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase</title><source>MEDLINE</source><source>Alma/SFX Local Collection</source><creator>Hiroko, Ishii-Ohba ; Hiroshi, Inano ; Bun-Ichi, Tamaoki</creator><creatorcontrib>Hiroko, Ishii-Ohba ; Hiroshi, Inano ; Bun-Ichi, Tamaoki</creatorcontrib><description>Through the treatment of rat testicular microsomes with sodium cholate, 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase (abbreviated as the 3β -hydroxysteroid dehydrogenase and isomerase, respectively) were solubilized, and then purified by DEAE and hydroxylapatite column chromatographies. The findings were as follows:
1. With this purification procedure, the 3β-hydroxysteroid dehydrogenase activity could not be separated from the isomerase.
2. For 3-oxo-4-ene-steroid formation from 3β-hydroxy-5-ene-steroids, NAD
+ was required as a cofactor. While the 3β-hydroxysteroid dehydrogenase required NAD
+, the isomerase also required NAD
+ or its reduced form, in contrast to the microbial enzyme.
3. On treatment of the purified enzyme with 5'-
p-fluorosulfonyl-benzoyladenosine (FSBA), both enzyme activities were markedly reduced.
4. The enzyme, affinity labeled with [adenine-8-
14C]FSBA, showed a mol. wt of 46.8 K.
5. During 4-androstenedione production from DHA, 5-androstenedione was detected as an intermediate.</description><identifier>ISSN: 0022-4731</identifier><identifier>DOI: 10.1016/0022-4731(86)90402-4</identifier><identifier>PMID: 2945972</identifier><identifier>CODEN: JSTBBK</identifier><language>eng</language><publisher>Oxford: Elsevier B.V</publisher><subject>3-Hydroxysteroid Dehydrogenases - isolation & purification ; Adenosine - analogs & derivatives ; Affinity Labels ; Analytical, structural and metabolic biochemistry ; Androstenedione - biosynthesis ; Animals ; Biological and medical sciences ; Coenzymes - metabolism ; Dehydroepiandrosterone - metabolism ; Enzymes and enzyme inhibitors ; Fundamental and applied biological sciences. Psychology ; General aspects, investigation methods ; Isomerases - isolation & purification ; Male ; Molecular Weight ; Rats ; Solubility ; Steroid Isomerases - isolation & purification ; Testis - enzymology</subject><ispartof>Journal of steroid biochemistry, 1986-10, Vol.25 (4), p.555-560</ispartof><rights>1986</rights><rights>1987 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c386t-8dc8fdc0e045d8371384d938ae66fff6e68d31f600051e69f0825cedc24e6f753</citedby><cites>FETCH-LOGICAL-c386t-8dc8fdc0e045d8371384d938ae66fff6e68d31f600051e69f0825cedc24e6f753</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>309,310,314,780,784,789,790,23928,23929,25138,27922,27923</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=8117680$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2945972$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hiroko, Ishii-Ohba</creatorcontrib><creatorcontrib>Hiroshi, Inano</creatorcontrib><creatorcontrib>Bun-Ichi, Tamaoki</creatorcontrib><title>Purification and properties of testicular 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase</title><title>Journal of steroid biochemistry</title><addtitle>J Steroid Biochem</addtitle><description>Through the treatment of rat testicular microsomes with sodium cholate, 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase (abbreviated as the 3β -hydroxysteroid dehydrogenase and isomerase, respectively) were solubilized, and then purified by DEAE and hydroxylapatite column chromatographies. The findings were as follows:
1. With this purification procedure, the 3β-hydroxysteroid dehydrogenase activity could not be separated from the isomerase.
2. For 3-oxo-4-ene-steroid formation from 3β-hydroxy-5-ene-steroids, NAD
+ was required as a cofactor. While the 3β-hydroxysteroid dehydrogenase required NAD
+, the isomerase also required NAD
+ or its reduced form, in contrast to the microbial enzyme.
3. On treatment of the purified enzyme with 5'-
p-fluorosulfonyl-benzoyladenosine (FSBA), both enzyme activities were markedly reduced.
4. The enzyme, affinity labeled with [adenine-8-
14C]FSBA, showed a mol. wt of 46.8 K.
5. During 4-androstenedione production from DHA, 5-androstenedione was detected as an intermediate.</description><subject>3-Hydroxysteroid Dehydrogenases - isolation & purification</subject><subject>Adenosine - analogs & derivatives</subject><subject>Affinity Labels</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Androstenedione - biosynthesis</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Coenzymes - metabolism</subject><subject>Dehydroepiandrosterone - metabolism</subject><subject>Enzymes and enzyme inhibitors</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>General aspects, investigation methods</subject><subject>Isomerases - isolation & purification</subject><subject>Male</subject><subject>Molecular Weight</subject><subject>Rats</subject><subject>Solubility</subject><subject>Steroid Isomerases - isolation & purification</subject><subject>Testis - enzymology</subject><issn>0022-4731</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1986</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kM1OxCAUhVloxnH0DTTpwhhdVKFQSjcmZuJfYqILXROEi6KdMkJrnNfyQXwm6cxklm4g3HPuCedD6IDgM4IJP8e4KHJWUXIi-GmNGU6vLTTejHfQbozvGJNasGKERkXNyroqxujjsQ_OOq0659tMtSabBz-H0DmImbdZB7Fzum9UyOjvT_62MMF_L_Iyhxby2EHwzmQGlvNXaFWEZchKZ8OZuehnEJKyh7ataiLsr-8Jer6-epre5vcPN3fTy_tcU8G7XBgtrNEYMCuNoBWhgpmaCgWcW2s5cGEosRxjXBLgtcWiKDUYXTDgtirpBB2vclOVzz4VkDMXNTSNasH3UVYpsqSkSka2MurgYwxg5Ty4mQoLSbAcuMoBoBwASsHlkqtkae1wnd-_zMBsltZQk3601lXUqrFBtdrFjU0QUnGBk-1iZYPE4stBkFE7aFMTF0B30nj3_z_-AHP-lyQ</recordid><startdate>19861001</startdate><enddate>19861001</enddate><creator>Hiroko, Ishii-Ohba</creator><creator>Hiroshi, Inano</creator><creator>Bun-Ichi, Tamaoki</creator><general>Elsevier B.V</general><general>Pergamon</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19861001</creationdate><title>Purification and properties of testicular 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase</title><author>Hiroko, Ishii-Ohba ; Hiroshi, Inano ; Bun-Ichi, Tamaoki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c386t-8dc8fdc0e045d8371384d938ae66fff6e68d31f600051e69f0825cedc24e6f753</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1986</creationdate><topic>3-Hydroxysteroid Dehydrogenases - isolation & purification</topic><topic>Adenosine - analogs & derivatives</topic><topic>Affinity Labels</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Androstenedione - biosynthesis</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Coenzymes - metabolism</topic><topic>Dehydroepiandrosterone - metabolism</topic><topic>Enzymes and enzyme inhibitors</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>General aspects, investigation methods</topic><topic>Isomerases - isolation & purification</topic><topic>Male</topic><topic>Molecular Weight</topic><topic>Rats</topic><topic>Solubility</topic><topic>Steroid Isomerases - isolation & purification</topic><topic>Testis - enzymology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hiroko, Ishii-Ohba</creatorcontrib><creatorcontrib>Hiroshi, Inano</creatorcontrib><creatorcontrib>Bun-Ichi, Tamaoki</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of steroid biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hiroko, Ishii-Ohba</au><au>Hiroshi, Inano</au><au>Bun-Ichi, Tamaoki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification and properties of testicular 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase</atitle><jtitle>Journal of steroid biochemistry</jtitle><addtitle>J Steroid Biochem</addtitle><date>1986-10-01</date><risdate>1986</risdate><volume>25</volume><issue>4</issue><spage>555</spage><epage>560</epage><pages>555-560</pages><issn>0022-4731</issn><coden>JSTBBK</coden><abstract>Through the treatment of rat testicular microsomes with sodium cholate, 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase (abbreviated as the 3β -hydroxysteroid dehydrogenase and isomerase, respectively) were solubilized, and then purified by DEAE and hydroxylapatite column chromatographies. The findings were as follows:
1. With this purification procedure, the 3β-hydroxysteroid dehydrogenase activity could not be separated from the isomerase.
2. For 3-oxo-4-ene-steroid formation from 3β-hydroxy-5-ene-steroids, NAD
+ was required as a cofactor. While the 3β-hydroxysteroid dehydrogenase required NAD
+, the isomerase also required NAD
+ or its reduced form, in contrast to the microbial enzyme.
3. On treatment of the purified enzyme with 5'-
p-fluorosulfonyl-benzoyladenosine (FSBA), both enzyme activities were markedly reduced.
4. The enzyme, affinity labeled with [adenine-8-
14C]FSBA, showed a mol. wt of 46.8 K.
5. During 4-androstenedione production from DHA, 5-androstenedione was detected as an intermediate.</abstract><cop>Oxford</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>2945972</pmid><doi>10.1016/0022-4731(86)90402-4</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0022-4731 |
| ispartof | Journal of steroid biochemistry, 1986-10, Vol.25 (4), p.555-560 |
| issn | 0022-4731 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_77135317 |
| source | MEDLINE; Alma/SFX Local Collection |
| subjects | 3-Hydroxysteroid Dehydrogenases - isolation & purification Adenosine - analogs & derivatives Affinity Labels Analytical, structural and metabolic biochemistry Androstenedione - biosynthesis Animals Biological and medical sciences Coenzymes - metabolism Dehydroepiandrosterone - metabolism Enzymes and enzyme inhibitors Fundamental and applied biological sciences. Psychology General aspects, investigation methods Isomerases - isolation & purification Male Molecular Weight Rats Solubility Steroid Isomerases - isolation & purification Testis - enzymology |
| title | Purification and properties of testicular 3β-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-14T06%3A57%3A03IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Purification%20and%20properties%20of%20testicular%203%CE%B2-hydroxy-5-ene-steroid%20dehydrogenase%20and%205-ene-4-ene%20isomerase&rft.jtitle=Journal%20of%20steroid%20biochemistry&rft.au=Hiroko,%20Ishii-Ohba&rft.date=1986-10-01&rft.volume=25&rft.issue=4&rft.spage=555&rft.epage=560&rft.pages=555-560&rft.issn=0022-4731&rft.coden=JSTBBK&rft_id=info:doi/10.1016/0022-4731(86)90402-4&rft_dat=%3Cproquest_cross%3E77135317%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=77135317&rft_id=info:pmid/2945972&rft_els_id=0022473186904024&rfr_iscdi=true |