Mechanistic and metabolic studies of sterol 24,25-double bond reduction in Manduca sexta
Larvae of Manduca sexta were used to obtain a cell-free sterol 24,25-reductase. From the midgut of fifth instar larvae fed a mixture of sitosterol and campesterol a microsome-bound 24,25-sterol reductase was prepared that transformed desmosterol (Km, 3 micromolar), lanosterol (Km,18 micromolar), and...
Gespeichert in:
| Veröffentlicht in: | Archives of insect biochemistry and physiology 1996, Vol.31 (1), p.1-22 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 22 |
|---|---|
| container_issue | 1 |
| container_start_page | 1 |
| container_title | Archives of insect biochemistry and physiology |
| container_volume | 31 |
| creator | Short, J.D. (Texas Tech University, Lubbock, TX.) Guo, D.A Svoboda, J.A Nes, W.D |
| description | Larvae of Manduca sexta were used to obtain a cell-free sterol 24,25-reductase. From the midgut of fifth instar larvae fed a mixture of sitosterol and campesterol a microsome-bound 24,25-sterol reductase was prepared that transformed desmosterol (Km, 3 micromolar), lanosterol (Km,18 micromolar), and cycloartenol (Km,33 micromolar) to cholesterol, 24,25-dihydrolanosterol, and cycloartanol, respectively. With desmosterol as substrate, the microsome-bound enzyme was found to incorporate tritium into cholesterol from 4S-tritium labelled NADPH. [24-2H]lanosterol was transformed by larvae to [24-2H]24,25-dihydrolanosterol (structure confirmed by mass spectroscopy (MS) and 1H-nuclear magnetic resonance spectroscopy. A rationally designed inhibitor of 24,25-reductase activity, 24(R,S),25-epiminolanosterol (IL), was assayed and found to be inhibitory with an I50 of 2 micromolar. IL was supplemented in the diet of M. sexta with either sitosterol or stigmasterol and found to inhibit development (I50, 60 ppm). The major sterol which accumulated in the IL-treated larvae was desmosterol, confirming the site of inhibition was reduction of the 24,25-bond. IL was converted to [2-3H]IL when fed to the larvae. [2-3H]lanosterol was recovered from fifth instar larvae and its structure confirmed by MS and radiochemical techniques |
| doi_str_mv | 10.1002/(SICI)1520-6327(1996)31:1<1::AID-ARCH1>3.0.CO;2-4 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_wiley</sourceid><recordid>TN_cdi_proquest_miscellaneous_17021284</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>17021284</sourcerecordid><originalsourceid>FETCH-LOGICAL-f3161-f920cb7387b06a0bed4143c9750388444f1d9fa66168c749b2a8a7cabba04a093</originalsourceid><addsrcrecordid>eNo9kUtv1DAURi0EEkPhD7DKCrUSGa4fseMBIQ0B2pE6jNRSQGyuHMeBlExc4kRt_32dpqq8uA-d-y2OCdEUlhSAvTs83xSbI5oxSCVn6pBqLY84XdEPdLVabz6n67PihH7kS1gWu_csFU_I4pF-ShaguE6FkOw5eRHCJQBoSfMF-bV19q_pmjA0NjFdlezdYErfxikMY9W4kPg6tq73bcLEW5allR_L1iWlj3TvqtEOje-Spku28X60JgnuZjAvybPatMG9eqgH5OLrl-_FSXq6O94U69O05lTStNYMbKl4rkqQBkpXCSq41SoDnudCiJpWujZSUplbJXTJTG6UNWVpQBjQ_IC8mXOvev9_dGHAfROsa1vTOT8GpAoYZbmI4NkMXjetu8Wrvtmb_hYp4CQYJ784GcPJGE5-kVOcXtSL93qRI2CxQ4ZiXsTQdA6NAt3NY6jp_6FUXGX489sxZr8h-yR_aNxG_vXM18aj-dM3AS_OtWRZ_B9-Bx6eiwI</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>17021284</pqid></control><display><type>article</type><title>Mechanistic and metabolic studies of sterol 24,25-double bond reduction in Manduca sexta</title><source>Wiley Online Library Journals Frontfile Complete</source><creator>Short, J.D. (Texas Tech University, Lubbock, TX.) ; Guo, D.A ; Svoboda, J.A ; Nes, W.D</creator><creatorcontrib>Short, J.D. (Texas Tech University, Lubbock, TX.) ; Guo, D.A ; Svoboda, J.A ; Nes, W.D</creatorcontrib><description>Larvae of Manduca sexta were used to obtain a cell-free sterol 24,25-reductase. From the midgut of fifth instar larvae fed a mixture of sitosterol and campesterol a microsome-bound 24,25-sterol reductase was prepared that transformed desmosterol (Km, 3 micromolar), lanosterol (Km,18 micromolar), and cycloartenol (Km,33 micromolar) to cholesterol, 24,25-dihydrolanosterol, and cycloartanol, respectively. With desmosterol as substrate, the microsome-bound enzyme was found to incorporate tritium into cholesterol from 4S-tritium labelled NADPH. [24-2H]lanosterol was transformed by larvae to [24-2H]24,25-dihydrolanosterol (structure confirmed by mass spectroscopy (MS) and 1H-nuclear magnetic resonance spectroscopy. A rationally designed inhibitor of 24,25-reductase activity, 24(R,S),25-epiminolanosterol (IL), was assayed and found to be inhibitory with an I50 of 2 micromolar. IL was supplemented in the diet of M. sexta with either sitosterol or stigmasterol and found to inhibit development (I50, 60 ppm). The major sterol which accumulated in the IL-treated larvae was desmosterol, confirming the site of inhibition was reduction of the 24,25-bond. IL was converted to [2-3H]IL when fed to the larvae. [2-3H]lanosterol was recovered from fifth instar larvae and its structure confirmed by MS and radiochemical techniques</description><identifier>ISSN: 0739-4462</identifier><identifier>EISSN: 1520-6327</identifier><identifier>DOI: 10.1002/(SICI)1520-6327(1996)31:1<1::AID-ARCH1>3.0.CO;2-4</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>ACTIVIDAD ENZIMATICA ; ACTIVITE ENZYMATIQUE ; cholesterol ; CRECIMIENTO ; CROISSANCE ; desmosterol ; DIETA ; ESTEROLES ; INHIBICION ; INHIBIDORES DE ENZIMAS ; INHIBITEUR D'ENZYME ; INHIBITION ; LARVAS ; LARVE ; Lepidoptera ; MANDUCA SEXTA ; METABOLISME DES STEROIDES ; METABOLISMO DE ESTEROIDES ; OXIDORREDUCTASAS ; OXYDOREDUCTASE ; REDUCCION ; REDUCTION ; REGIME ALIMENTAIRE ; sitosterol ; Sphingidae ; STEROL ; tobacco hornworm</subject><ispartof>Archives of insect biochemistry and physiology, 1996, Vol.31 (1), p.1-22</ispartof><rights>Copyright © 1996 Wiley‐Liss, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2F%28SICI%291520-6327%281996%2931%3A1%3C1%3A%3AAID-ARCH1%3E3.0.CO%3B2-4$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2F%28SICI%291520-6327%281996%2931%3A1%3C1%3A%3AAID-ARCH1%3E3.0.CO%3B2-4$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,777,781,1412,4010,27904,27905,27906,45555,45556</link.rule.ids></links><search><creatorcontrib>Short, J.D. (Texas Tech University, Lubbock, TX.)</creatorcontrib><creatorcontrib>Guo, D.A</creatorcontrib><creatorcontrib>Svoboda, J.A</creatorcontrib><creatorcontrib>Nes, W.D</creatorcontrib><title>Mechanistic and metabolic studies of sterol 24,25-double bond reduction in Manduca sexta</title><title>Archives of insect biochemistry and physiology</title><addtitle>Arch. Insect Biochem. Physiol</addtitle><description>Larvae of Manduca sexta were used to obtain a cell-free sterol 24,25-reductase. From the midgut of fifth instar larvae fed a mixture of sitosterol and campesterol a microsome-bound 24,25-sterol reductase was prepared that transformed desmosterol (Km, 3 micromolar), lanosterol (Km,18 micromolar), and cycloartenol (Km,33 micromolar) to cholesterol, 24,25-dihydrolanosterol, and cycloartanol, respectively. With desmosterol as substrate, the microsome-bound enzyme was found to incorporate tritium into cholesterol from 4S-tritium labelled NADPH. [24-2H]lanosterol was transformed by larvae to [24-2H]24,25-dihydrolanosterol (structure confirmed by mass spectroscopy (MS) and 1H-nuclear magnetic resonance spectroscopy. A rationally designed inhibitor of 24,25-reductase activity, 24(R,S),25-epiminolanosterol (IL), was assayed and found to be inhibitory with an I50 of 2 micromolar. IL was supplemented in the diet of M. sexta with either sitosterol or stigmasterol and found to inhibit development (I50, 60 ppm). The major sterol which accumulated in the IL-treated larvae was desmosterol, confirming the site of inhibition was reduction of the 24,25-bond. IL was converted to [2-3H]IL when fed to the larvae. [2-3H]lanosterol was recovered from fifth instar larvae and its structure confirmed by MS and radiochemical techniques</description><subject>ACTIVIDAD ENZIMATICA</subject><subject>ACTIVITE ENZYMATIQUE</subject><subject>cholesterol</subject><subject>CRECIMIENTO</subject><subject>CROISSANCE</subject><subject>desmosterol</subject><subject>DIETA</subject><subject>ESTEROLES</subject><subject>INHIBICION</subject><subject>INHIBIDORES DE ENZIMAS</subject><subject>INHIBITEUR D'ENZYME</subject><subject>INHIBITION</subject><subject>LARVAS</subject><subject>LARVE</subject><subject>Lepidoptera</subject><subject>MANDUCA SEXTA</subject><subject>METABOLISME DES STEROIDES</subject><subject>METABOLISMO DE ESTEROIDES</subject><subject>OXIDORREDUCTASAS</subject><subject>OXYDOREDUCTASE</subject><subject>REDUCCION</subject><subject>REDUCTION</subject><subject>REGIME ALIMENTAIRE</subject><subject>sitosterol</subject><subject>Sphingidae</subject><subject>STEROL</subject><subject>tobacco hornworm</subject><issn>0739-4462</issn><issn>1520-6327</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><recordid>eNo9kUtv1DAURi0EEkPhD7DKCrUSGa4fseMBIQ0B2pE6jNRSQGyuHMeBlExc4kRt_32dpqq8uA-d-y2OCdEUlhSAvTs83xSbI5oxSCVn6pBqLY84XdEPdLVabz6n67PihH7kS1gWu_csFU_I4pF-ShaguE6FkOw5eRHCJQBoSfMF-bV19q_pmjA0NjFdlezdYErfxikMY9W4kPg6tq73bcLEW5allR_L1iWlj3TvqtEOje-Spku28X60JgnuZjAvybPatMG9eqgH5OLrl-_FSXq6O94U69O05lTStNYMbKl4rkqQBkpXCSq41SoDnudCiJpWujZSUplbJXTJTG6UNWVpQBjQ_IC8mXOvev9_dGHAfROsa1vTOT8GpAoYZbmI4NkMXjetu8Wrvtmb_hYp4CQYJ784GcPJGE5-kVOcXtSL93qRI2CxQ4ZiXsTQdA6NAt3NY6jp_6FUXGX489sxZr8h-yR_aNxG_vXM18aj-dM3AS_OtWRZ_B9-Bx6eiwI</recordid><startdate>1996</startdate><enddate>1996</enddate><creator>Short, J.D. (Texas Tech University, Lubbock, TX.)</creator><creator>Guo, D.A</creator><creator>Svoboda, J.A</creator><creator>Nes, W.D</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><scope>FBQ</scope><scope>BSCLL</scope><scope>7SS</scope></search><sort><creationdate>1996</creationdate><title>Mechanistic and metabolic studies of sterol 24,25-double bond reduction in Manduca sexta</title><author>Short, J.D. (Texas Tech University, Lubbock, TX.) ; Guo, D.A ; Svoboda, J.A ; Nes, W.D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-f3161-f920cb7387b06a0bed4143c9750388444f1d9fa66168c749b2a8a7cabba04a093</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>ACTIVIDAD ENZIMATICA</topic><topic>ACTIVITE ENZYMATIQUE</topic><topic>cholesterol</topic><topic>CRECIMIENTO</topic><topic>CROISSANCE</topic><topic>desmosterol</topic><topic>DIETA</topic><topic>ESTEROLES</topic><topic>INHIBICION</topic><topic>INHIBIDORES DE ENZIMAS</topic><topic>INHIBITEUR D'ENZYME</topic><topic>INHIBITION</topic><topic>LARVAS</topic><topic>LARVE</topic><topic>Lepidoptera</topic><topic>MANDUCA SEXTA</topic><topic>METABOLISME DES STEROIDES</topic><topic>METABOLISMO DE ESTEROIDES</topic><topic>OXIDORREDUCTASAS</topic><topic>OXYDOREDUCTASE</topic><topic>REDUCCION</topic><topic>REDUCTION</topic><topic>REGIME ALIMENTAIRE</topic><topic>sitosterol</topic><topic>Sphingidae</topic><topic>STEROL</topic><topic>tobacco hornworm</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Short, J.D. (Texas Tech University, Lubbock, TX.)</creatorcontrib><creatorcontrib>Guo, D.A</creatorcontrib><creatorcontrib>Svoboda, J.A</creatorcontrib><creatorcontrib>Nes, W.D</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Entomology Abstracts (Full archive)</collection><jtitle>Archives of insect biochemistry and physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Short, J.D. (Texas Tech University, Lubbock, TX.)</au><au>Guo, D.A</au><au>Svoboda, J.A</au><au>Nes, W.D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mechanistic and metabolic studies of sterol 24,25-double bond reduction in Manduca sexta</atitle><jtitle>Archives of insect biochemistry and physiology</jtitle><addtitle>Arch. Insect Biochem. Physiol</addtitle><date>1996</date><risdate>1996</risdate><volume>31</volume><issue>1</issue><spage>1</spage><epage>22</epage><pages>1-22</pages><issn>0739-4462</issn><eissn>1520-6327</eissn><abstract>Larvae of Manduca sexta were used to obtain a cell-free sterol 24,25-reductase. From the midgut of fifth instar larvae fed a mixture of sitosterol and campesterol a microsome-bound 24,25-sterol reductase was prepared that transformed desmosterol (Km, 3 micromolar), lanosterol (Km,18 micromolar), and cycloartenol (Km,33 micromolar) to cholesterol, 24,25-dihydrolanosterol, and cycloartanol, respectively. With desmosterol as substrate, the microsome-bound enzyme was found to incorporate tritium into cholesterol from 4S-tritium labelled NADPH. [24-2H]lanosterol was transformed by larvae to [24-2H]24,25-dihydrolanosterol (structure confirmed by mass spectroscopy (MS) and 1H-nuclear magnetic resonance spectroscopy. A rationally designed inhibitor of 24,25-reductase activity, 24(R,S),25-epiminolanosterol (IL), was assayed and found to be inhibitory with an I50 of 2 micromolar. IL was supplemented in the diet of M. sexta with either sitosterol or stigmasterol and found to inhibit development (I50, 60 ppm). The major sterol which accumulated in the IL-treated larvae was desmosterol, confirming the site of inhibition was reduction of the 24,25-bond. IL was converted to [2-3H]IL when fed to the larvae. [2-3H]lanosterol was recovered from fifth instar larvae and its structure confirmed by MS and radiochemical techniques</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><doi>10.1002/(SICI)1520-6327(1996)31:1<1::AID-ARCH1>3.0.CO;2-4</doi><tpages>22</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0739-4462 |
| ispartof | Archives of insect biochemistry and physiology, 1996, Vol.31 (1), p.1-22 |
| issn | 0739-4462 1520-6327 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_17021284 |
| source | Wiley Online Library Journals Frontfile Complete |
| subjects | ACTIVIDAD ENZIMATICA ACTIVITE ENZYMATIQUE cholesterol CRECIMIENTO CROISSANCE desmosterol DIETA ESTEROLES INHIBICION INHIBIDORES DE ENZIMAS INHIBITEUR D'ENZYME INHIBITION LARVAS LARVE Lepidoptera MANDUCA SEXTA METABOLISME DES STEROIDES METABOLISMO DE ESTEROIDES OXIDORREDUCTASAS OXYDOREDUCTASE REDUCCION REDUCTION REGIME ALIMENTAIRE sitosterol Sphingidae STEROL tobacco hornworm |
| title | Mechanistic and metabolic studies of sterol 24,25-double bond reduction in Manduca sexta |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-20T04%3A33%3A06IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_wiley&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Mechanistic%20and%20metabolic%20studies%20of%20sterol%2024,25-double%20bond%20reduction%20in%20Manduca%20sexta&rft.jtitle=Archives%20of%20insect%20biochemistry%20and%20physiology&rft.au=Short,%20J.D.%20(Texas%20Tech%20University,%20Lubbock,%20TX.)&rft.date=1996&rft.volume=31&rft.issue=1&rft.spage=1&rft.epage=22&rft.pages=1-22&rft.issn=0739-4462&rft.eissn=1520-6327&rft_id=info:doi/10.1002/(SICI)1520-6327(1996)31:1%3C1::AID-ARCH1%3E3.0.CO;2-4&rft_dat=%3Cproquest_wiley%3E17021284%3C/proquest_wiley%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=17021284&rft_id=info:pmid/&rfr_iscdi=true |