Enzymatic properties of vesicle-reconstituted human cytochrome P450SCC (CYP11A1) differences in functioning of the mitochondrial electron-transfer chain using human and bovine adrenodoxin and activation by cardiolipin

The recently reported heterologous expression and purification of both human cytochrome P450SCC and adrenodoxin [Woods, S.T., Sadleir, J., Downs, T., Triantopoulos, T., Haedlam, M.J. & Tuckey, R.C. (1998) Arch. Biochem. Biophys. 353, 109-115] has enabled us to perform studies with the membrane-r...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:European journal of biochemistry 1999-03, Vol.260 (3), p.768-773
Hauptverfasser: Kisselev, P, Tuckey, R C, Woods, S T, Triantopoulos, T, Schwarz, D
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 773
container_issue 3
container_start_page 768
container_title European journal of biochemistry
container_volume 260
creator Kisselev, P
Tuckey, R C
Woods, S T
Triantopoulos, T
Schwarz, D
description The recently reported heterologous expression and purification of both human cytochrome P450SCC and adrenodoxin [Woods, S.T., Sadleir, J., Downs, T., Triantopoulos, T., Haedlam, M.J. & Tuckey, R.C. (1998) Arch. Biochem. Biophys. 353, 109-115] has enabled us to perform studies with the membrane-reconstituted human enzymes to better understand the side-chain cleavage reaction in humans. Human P450SCC was successfully reconstituted into dioleoylphosphatidylcholine vesicles with and without cardiolipin and its enzymatic properties characterized in the membrane-bound state. Enhancement of the P450SCC activity and significant activation by cardiolipin were observed when human adrenodoxin instead of bovine adrenodoxin was used as electron donor. In the absence of cardiolipin, Km for cholesterol was decreased twice in the case of human adrenodoxin indicating enhanced cholesterol binding. On the other hand, in the presence of cardiolipin in the membrane both Km and V for cholesterol were decreased with human adrenodoxin as electron donor. Kinetic analysis of the interaction between human P450SCC and its redox partners provided evidence for enhanced binding of the human electron donor to human P450SCC indicated by both an increased V and decreased Kd for human adrenodoxin compared with the values with bovine adrenodoxin. Because no similar effects were observed in Tween 20 micelles, these results suggest that the phospholipid membrane may play an important role in the interaction of human adrenodoxin with human P450SCC.
doi_str_mv 10.1046/j.1432-1327.1999.00206.x
format Article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_69665804</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>69665804</sourcerecordid><originalsourceid>FETCH-LOGICAL-c256t-f3cbe677949a946af80347dc9c79f8c74a6d75aeb0febb9a4e828c5b87f1bb5f3</originalsourceid><addsrcrecordid>eNpNkU-P1CAYh4nRuLOrX8FwMnpohf6h5bhpVtdkEzdRD54I0BeHSQsj0MmM39RvI7V78ESA9_n9kvdBCFNSUtKwD4eSNnVV0LrqSso5LwmpCCvPz9Bu-yB1_RztCKFNUfGWXaHrGA-EEMZZ9xJdUZIn8m2H_ty535dZJqvxMfgjhGQhYm_wCaLVExQBtHcx2bQkGPF-maXD-pK83gc_A35sWvJ1GPC74ccjpbf0PR6tMRDA6ZxjHTaL08l6Z93PNTbtAc92xb0bg5UThgl0Ct4VKUgXM4r1XmZwiSuyFUo3YuVP1gGWY872oz_b7Vnm9JNcG7C6YC3DaP1kj9a9Qi-MnCK8fjpv0PePd9-G--Lhy6fPw-1DoauWpcLUWgHrOt5wyRsmTU_qphs11x03ve4aycaulaCIAaW4bKCvet2qvjNUqdbUN-jtlpv392uBmMRso4Zpkg78EkVeOWt70uTBfhvUwccYwIhjsLMMF0GJWLWKg1jtiVWrWLWKf1rFOaNvnjoWNcP4H7h5rP8ConOlXw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>69665804</pqid></control><display><type>article</type><title>Enzymatic properties of vesicle-reconstituted human cytochrome P450SCC (CYP11A1) differences in functioning of the mitochondrial electron-transfer chain using human and bovine adrenodoxin and activation by cardiolipin</title><source>MEDLINE</source><source>Access via Wiley Online Library</source><source>Alma/SFX Local Collection</source><creator>Kisselev, P ; Tuckey, R C ; Woods, S T ; Triantopoulos, T ; Schwarz, D</creator><creatorcontrib>Kisselev, P ; Tuckey, R C ; Woods, S T ; Triantopoulos, T ; Schwarz, D</creatorcontrib><description>The recently reported heterologous expression and purification of both human cytochrome P450SCC and adrenodoxin [Woods, S.T., Sadleir, J., Downs, T., Triantopoulos, T., Haedlam, M.J. &amp; Tuckey, R.C. (1998) Arch. Biochem. Biophys. 353, 109-115] has enabled us to perform studies with the membrane-reconstituted human enzymes to better understand the side-chain cleavage reaction in humans. Human P450SCC was successfully reconstituted into dioleoylphosphatidylcholine vesicles with and without cardiolipin and its enzymatic properties characterized in the membrane-bound state. Enhancement of the P450SCC activity and significant activation by cardiolipin were observed when human adrenodoxin instead of bovine adrenodoxin was used as electron donor. In the absence of cardiolipin, Km for cholesterol was decreased twice in the case of human adrenodoxin indicating enhanced cholesterol binding. On the other hand, in the presence of cardiolipin in the membrane both Km and V for cholesterol were decreased with human adrenodoxin as electron donor. Kinetic analysis of the interaction between human P450SCC and its redox partners provided evidence for enhanced binding of the human electron donor to human P450SCC indicated by both an increased V and decreased Kd for human adrenodoxin compared with the values with bovine adrenodoxin. Because no similar effects were observed in Tween 20 micelles, these results suggest that the phospholipid membrane may play an important role in the interaction of human adrenodoxin with human P450SCC.</description><identifier>ISSN: 0014-2956</identifier><identifier>EISSN: 1432-1033</identifier><identifier>DOI: 10.1046/j.1432-1327.1999.00206.x</identifier><identifier>PMID: 10103006</identifier><language>eng</language><publisher>England</publisher><subject>Adrenodoxin - pharmacology ; Animals ; Binding Sites ; Cardiolipins - pharmacology ; Cattle ; Cholesterol - metabolism ; Cholesterol Side-Chain Cleavage Enzyme - isolation &amp; purification ; Cholesterol Side-Chain Cleavage Enzyme - metabolism ; Electron Transport - drug effects ; Enzyme Activation ; Humans ; Kinetics ; Micelles ; Mitochondria - drug effects ; Mitochondria - enzymology ; Mitochondria - metabolism ; Oxidation-Reduction ; Phospholipids - metabolism ; Polysorbates - pharmacology ; Species Specificity</subject><ispartof>European journal of biochemistry, 1999-03, Vol.260 (3), p.768-773</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c256t-f3cbe677949a946af80347dc9c79f8c74a6d75aeb0febb9a4e828c5b87f1bb5f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,27929,27930</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10103006$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kisselev, P</creatorcontrib><creatorcontrib>Tuckey, R C</creatorcontrib><creatorcontrib>Woods, S T</creatorcontrib><creatorcontrib>Triantopoulos, T</creatorcontrib><creatorcontrib>Schwarz, D</creatorcontrib><title>Enzymatic properties of vesicle-reconstituted human cytochrome P450SCC (CYP11A1) differences in functioning of the mitochondrial electron-transfer chain using human and bovine adrenodoxin and activation by cardiolipin</title><title>European journal of biochemistry</title><addtitle>Eur J Biochem</addtitle><description>The recently reported heterologous expression and purification of both human cytochrome P450SCC and adrenodoxin [Woods, S.T., Sadleir, J., Downs, T., Triantopoulos, T., Haedlam, M.J. &amp; Tuckey, R.C. (1998) Arch. Biochem. Biophys. 353, 109-115] has enabled us to perform studies with the membrane-reconstituted human enzymes to better understand the side-chain cleavage reaction in humans. Human P450SCC was successfully reconstituted into dioleoylphosphatidylcholine vesicles with and without cardiolipin and its enzymatic properties characterized in the membrane-bound state. Enhancement of the P450SCC activity and significant activation by cardiolipin were observed when human adrenodoxin instead of bovine adrenodoxin was used as electron donor. In the absence of cardiolipin, Km for cholesterol was decreased twice in the case of human adrenodoxin indicating enhanced cholesterol binding. On the other hand, in the presence of cardiolipin in the membrane both Km and V for cholesterol were decreased with human adrenodoxin as electron donor. Kinetic analysis of the interaction between human P450SCC and its redox partners provided evidence for enhanced binding of the human electron donor to human P450SCC indicated by both an increased V and decreased Kd for human adrenodoxin compared with the values with bovine adrenodoxin. Because no similar effects were observed in Tween 20 micelles, these results suggest that the phospholipid membrane may play an important role in the interaction of human adrenodoxin with human P450SCC.</description><subject>Adrenodoxin - pharmacology</subject><subject>Animals</subject><subject>Binding Sites</subject><subject>Cardiolipins - pharmacology</subject><subject>Cattle</subject><subject>Cholesterol - metabolism</subject><subject>Cholesterol Side-Chain Cleavage Enzyme - isolation &amp; purification</subject><subject>Cholesterol Side-Chain Cleavage Enzyme - metabolism</subject><subject>Electron Transport - drug effects</subject><subject>Enzyme Activation</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Micelles</subject><subject>Mitochondria - drug effects</subject><subject>Mitochondria - enzymology</subject><subject>Mitochondria - metabolism</subject><subject>Oxidation-Reduction</subject><subject>Phospholipids - metabolism</subject><subject>Polysorbates - pharmacology</subject><subject>Species Specificity</subject><issn>0014-2956</issn><issn>1432-1033</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpNkU-P1CAYh4nRuLOrX8FwMnpohf6h5bhpVtdkEzdRD54I0BeHSQsj0MmM39RvI7V78ESA9_n9kvdBCFNSUtKwD4eSNnVV0LrqSso5LwmpCCvPz9Bu-yB1_RztCKFNUfGWXaHrGA-EEMZZ9xJdUZIn8m2H_ty535dZJqvxMfgjhGQhYm_wCaLVExQBtHcx2bQkGPF-maXD-pK83gc_A35sWvJ1GPC74ccjpbf0PR6tMRDA6ZxjHTaL08l6Z93PNTbtAc92xb0bg5UThgl0Ct4VKUgXM4r1XmZwiSuyFUo3YuVP1gGWY872oz_b7Vnm9JNcG7C6YC3DaP1kj9a9Qi-MnCK8fjpv0PePd9-G--Lhy6fPw-1DoauWpcLUWgHrOt5wyRsmTU_qphs11x03ve4aycaulaCIAaW4bKCvet2qvjNUqdbUN-jtlpv392uBmMRso4Zpkg78EkVeOWt70uTBfhvUwccYwIhjsLMMF0GJWLWKg1jtiVWrWLWKf1rFOaNvnjoWNcP4H7h5rP8ConOlXw</recordid><startdate>19990301</startdate><enddate>19990301</enddate><creator>Kisselev, P</creator><creator>Tuckey, R C</creator><creator>Woods, S T</creator><creator>Triantopoulos, T</creator><creator>Schwarz, D</creator><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>19990301</creationdate><title>Enzymatic properties of vesicle-reconstituted human cytochrome P450SCC (CYP11A1) differences in functioning of the mitochondrial electron-transfer chain using human and bovine adrenodoxin and activation by cardiolipin</title><author>Kisselev, P ; Tuckey, R C ; Woods, S T ; Triantopoulos, T ; Schwarz, D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c256t-f3cbe677949a946af80347dc9c79f8c74a6d75aeb0febb9a4e828c5b87f1bb5f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>Adrenodoxin - pharmacology</topic><topic>Animals</topic><topic>Binding Sites</topic><topic>Cardiolipins - pharmacology</topic><topic>Cattle</topic><topic>Cholesterol - metabolism</topic><topic>Cholesterol Side-Chain Cleavage Enzyme - isolation &amp; purification</topic><topic>Cholesterol Side-Chain Cleavage Enzyme - metabolism</topic><topic>Electron Transport - drug effects</topic><topic>Enzyme Activation</topic><topic>Humans</topic><topic>Kinetics</topic><topic>Micelles</topic><topic>Mitochondria - drug effects</topic><topic>Mitochondria - enzymology</topic><topic>Mitochondria - metabolism</topic><topic>Oxidation-Reduction</topic><topic>Phospholipids - metabolism</topic><topic>Polysorbates - pharmacology</topic><topic>Species Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kisselev, P</creatorcontrib><creatorcontrib>Tuckey, R C</creatorcontrib><creatorcontrib>Woods, S T</creatorcontrib><creatorcontrib>Triantopoulos, T</creatorcontrib><creatorcontrib>Schwarz, D</creatorcontrib><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>European journal of biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kisselev, P</au><au>Tuckey, R C</au><au>Woods, S T</au><au>Triantopoulos, T</au><au>Schwarz, D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enzymatic properties of vesicle-reconstituted human cytochrome P450SCC (CYP11A1) differences in functioning of the mitochondrial electron-transfer chain using human and bovine adrenodoxin and activation by cardiolipin</atitle><jtitle>European journal of biochemistry</jtitle><addtitle>Eur J Biochem</addtitle><date>1999-03-01</date><risdate>1999</risdate><volume>260</volume><issue>3</issue><spage>768</spage><epage>773</epage><pages>768-773</pages><issn>0014-2956</issn><eissn>1432-1033</eissn><abstract>The recently reported heterologous expression and purification of both human cytochrome P450SCC and adrenodoxin [Woods, S.T., Sadleir, J., Downs, T., Triantopoulos, T., Haedlam, M.J. &amp; Tuckey, R.C. (1998) Arch. Biochem. Biophys. 353, 109-115] has enabled us to perform studies with the membrane-reconstituted human enzymes to better understand the side-chain cleavage reaction in humans. Human P450SCC was successfully reconstituted into dioleoylphosphatidylcholine vesicles with and without cardiolipin and its enzymatic properties characterized in the membrane-bound state. Enhancement of the P450SCC activity and significant activation by cardiolipin were observed when human adrenodoxin instead of bovine adrenodoxin was used as electron donor. In the absence of cardiolipin, Km for cholesterol was decreased twice in the case of human adrenodoxin indicating enhanced cholesterol binding. On the other hand, in the presence of cardiolipin in the membrane both Km and V for cholesterol were decreased with human adrenodoxin as electron donor. Kinetic analysis of the interaction between human P450SCC and its redox partners provided evidence for enhanced binding of the human electron donor to human P450SCC indicated by both an increased V and decreased Kd for human adrenodoxin compared with the values with bovine adrenodoxin. Because no similar effects were observed in Tween 20 micelles, these results suggest that the phospholipid membrane may play an important role in the interaction of human adrenodoxin with human P450SCC.</abstract><cop>England</cop><pmid>10103006</pmid><doi>10.1046/j.1432-1327.1999.00206.x</doi><tpages>6</tpages></addata></record>
fulltext fulltext
identifier ISSN: 0014-2956
ispartof European journal of biochemistry, 1999-03, Vol.260 (3), p.768-773
issn 0014-2956
1432-1033
language eng
recordid cdi_proquest_miscellaneous_69665804
source MEDLINE; Access via Wiley Online Library; Alma/SFX Local Collection
subjects Adrenodoxin - pharmacology
Animals
Binding Sites
Cardiolipins - pharmacology
Cattle
Cholesterol - metabolism
Cholesterol Side-Chain Cleavage Enzyme - isolation & purification
Cholesterol Side-Chain Cleavage Enzyme - metabolism
Electron Transport - drug effects
Enzyme Activation
Humans
Kinetics
Micelles
Mitochondria - drug effects
Mitochondria - enzymology
Mitochondria - metabolism
Oxidation-Reduction
Phospholipids - metabolism
Polysorbates - pharmacology
Species Specificity
title Enzymatic properties of vesicle-reconstituted human cytochrome P450SCC (CYP11A1) differences in functioning of the mitochondrial electron-transfer chain using human and bovine adrenodoxin and activation by cardiolipin
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-16T06%3A27%3A51IST&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=Enzymatic%20properties%20of%20vesicle-reconstituted%20human%20cytochrome%20P450SCC%20(CYP11A1)%20differences%20in%20functioning%20of%20the%20mitochondrial%20electron-transfer%20chain%20using%20human%20and%20bovine%20adrenodoxin%20and%20activation%20by%20cardiolipin&rft.jtitle=European%20journal%20of%20biochemistry&rft.au=Kisselev,%20P&rft.date=1999-03-01&rft.volume=260&rft.issue=3&rft.spage=768&rft.epage=773&rft.pages=768-773&rft.issn=0014-2956&rft.eissn=1432-1033&rft_id=info:doi/10.1046/j.1432-1327.1999.00206.x&rft_dat=%3Cproquest_cross%3E69665804%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=69665804&rft_id=info:pmid/10103006&rfr_iscdi=true