Structure of a Substrate Complex of Mammalian Cytochrome P450 2C5 at 2.3 Å Resolution:  Evidence for Multiple Substrate Binding Modes

The structure of rabbit microsomal cytochrome P450 2C5/3LVdH complexed with a substrate, 4-methyl-N-methyl-N-(2-phenyl-2H-pyrazol-3-yl)benzenesulfonamide (DMZ), was determined by X-ray crystallography to 2.3 Å resolution. Substrate docking studies and electron density maps indicate that DMZ binds to...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Biochemistry (Easton) 2003-06, Vol.42 (21), p.6370-6379
Hauptverfasser: Wester, Michael R, Johnson, Eric F, Marques-Soares, Cristina, Dansette, Patrick M, Mansuy, Daniel, Stout, C. David
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 6379
container_issue 21
container_start_page 6370
container_title Biochemistry (Easton)
container_volume 42
creator Wester, Michael R
Johnson, Eric F
Marques-Soares, Cristina
Dansette, Patrick M
Mansuy, Daniel
Stout, C. David
description The structure of rabbit microsomal cytochrome P450 2C5/3LVdH complexed with a substrate, 4-methyl-N-methyl-N-(2-phenyl-2H-pyrazol-3-yl)benzenesulfonamide (DMZ), was determined by X-ray crystallography to 2.3 Å resolution. Substrate docking studies and electron density maps indicate that DMZ binds to the enzyme in two antiparallel orientations of the long axis of the substrate. One orientation places the principal site of hydroxylation, the 4-methyl group, 4.4 Å from the heme Fe, whereas the alternate conformation positions the second, infrequent site of hydroxylation at >5.9 Å from the heme Fe. Comparison of this structure to that obtained previously for the enzyme indicates that the protein closes around the substrate and prevents open access of water from bulk solvent to the heme Fe. This reflects a ∼1.5 Å movement of the F and G helices relative to helix I. The present structure provides a complete model for the protein from residues 27−488 and defines two new helices F‘ and G‘. The G‘ helix is likely to contribute to interactions of the enzyme with membranes. The relatively large active site, as compared to the volume occupied by the substrate, and the flexibility of the enzyme are likely to underlie the capacity of drug-metabolizing enzymes to metabolize structurally diverse substrates of different sizes.
doi_str_mv 10.1021/bi0273922
format Article
fullrecord <record><control><sourceid>acs_hal_p</sourceid><recordid>TN_cdi_hal_primary_oai_HAL_hal_00068735v1</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>a52673040</sourcerecordid><originalsourceid>FETCH-LOGICAL-a364t-dd2834be0e064bcdbcf5df0f9b3096c77862e5adeab87ce410dcdd3e01aab82d3</originalsourceid><addsrcrecordid>eNptkM9O3DAQh62qqCy0h75A5UsPPQT8J4kTbhBRKNoViKUSN8uxJ8WQxCvbQXDjgtQ-V9-EJyGrRQsHTqP5zTcz0ofQV0p2KGF0t7aECV4y9gFNaMZIkpZl9hFNCCF5wsqcbKKtEK7HNiUi_YQ2KRO5YLSYoL_z6AcdBw_YNVjh-VCH6FUEXLlu0cLdMp6prlOtVT2u7qPTV951gM_SjGBWZVhFzHY4_v-IzyG4dojW9XtPD__w4a010GvAjfN4NrTRjgfffDiwvbH9HzxzBsJntNGoNsCXl7qNfv88vKiOk-np0a9qf5oonqcxMYYVPK2BAMnTWptaN5lpSFPWnJS5FqLIGWTKgKoLoSGlxGhjOBCqxoQZvo1-rO5eqVYuvO2Uv5dOWXm8P5XLbCmtEDy7pa-s9i4ED816gRK5NC_X5kf224pdDHUH5pV8UT0CyQqwIcLdeq78jcwFF5m8OJvLg0s6uzznJ1KM_PcVr3SQ127w_ajlncfPwwKaRA</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype></control><display><type>article</type><title>Structure of a Substrate Complex of Mammalian Cytochrome P450 2C5 at 2.3 Å Resolution:  Evidence for Multiple Substrate Binding Modes</title><source>American Chemical Society</source><source>MEDLINE</source><creator>Wester, Michael R ; Johnson, Eric F ; Marques-Soares, Cristina ; Dansette, Patrick M ; Mansuy, Daniel ; Stout, C. David</creator><creatorcontrib>Wester, Michael R ; Johnson, Eric F ; Marques-Soares, Cristina ; Dansette, Patrick M ; Mansuy, Daniel ; Stout, C. David</creatorcontrib><description>The structure of rabbit microsomal cytochrome P450 2C5/3LVdH complexed with a substrate, 4-methyl-N-methyl-N-(2-phenyl-2H-pyrazol-3-yl)benzenesulfonamide (DMZ), was determined by X-ray crystallography to 2.3 Å resolution. Substrate docking studies and electron density maps indicate that DMZ binds to the enzyme in two antiparallel orientations of the long axis of the substrate. One orientation places the principal site of hydroxylation, the 4-methyl group, 4.4 Å from the heme Fe, whereas the alternate conformation positions the second, infrequent site of hydroxylation at &gt;5.9 Å from the heme Fe. Comparison of this structure to that obtained previously for the enzyme indicates that the protein closes around the substrate and prevents open access of water from bulk solvent to the heme Fe. This reflects a ∼1.5 Å movement of the F and G helices relative to helix I. The present structure provides a complete model for the protein from residues 27−488 and defines two new helices F‘ and G‘. The G‘ helix is likely to contribute to interactions of the enzyme with membranes. The relatively large active site, as compared to the volume occupied by the substrate, and the flexibility of the enzyme are likely to underlie the capacity of drug-metabolizing enzymes to metabolize structurally diverse substrates of different sizes.</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi0273922</identifier><identifier>PMID: 12767218</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Biochemistry, Molecular Biology ; Crystallography, X-Ray ; Cytochrome P-450 Enzyme System - chemistry ; Cytochrome P450 Family 2 ; Electrons ; Heme - chemistry ; Humans ; Imidazoles - chemistry ; Iron - chemistry ; Life Sciences ; Models, Molecular ; Protein Binding ; Protein Conformation ; Protein Structure, Tertiary ; Steroid 21-Hydroxylase - chemistry ; Toxicology ; Water - chemistry</subject><ispartof>Biochemistry (Easton), 2003-06, Vol.42 (21), p.6370-6379</ispartof><rights>Copyright © 2003 American Chemical Society</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a364t-dd2834be0e064bcdbcf5df0f9b3096c77862e5adeab87ce410dcdd3e01aab82d3</citedby><cites>FETCH-LOGICAL-a364t-dd2834be0e064bcdbcf5df0f9b3096c77862e5adeab87ce410dcdd3e01aab82d3</cites><orcidid>0000-0003-0024-0023 ; 0000-0002-3694-3348</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/bi0273922$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bi0273922$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>230,314,780,784,885,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12767218$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-00068735$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Wester, Michael R</creatorcontrib><creatorcontrib>Johnson, Eric F</creatorcontrib><creatorcontrib>Marques-Soares, Cristina</creatorcontrib><creatorcontrib>Dansette, Patrick M</creatorcontrib><creatorcontrib>Mansuy, Daniel</creatorcontrib><creatorcontrib>Stout, C. David</creatorcontrib><title>Structure of a Substrate Complex of Mammalian Cytochrome P450 2C5 at 2.3 Å Resolution:  Evidence for Multiple Substrate Binding Modes</title><title>Biochemistry (Easton)</title><addtitle>Biochemistry</addtitle><description>The structure of rabbit microsomal cytochrome P450 2C5/3LVdH complexed with a substrate, 4-methyl-N-methyl-N-(2-phenyl-2H-pyrazol-3-yl)benzenesulfonamide (DMZ), was determined by X-ray crystallography to 2.3 Å resolution. Substrate docking studies and electron density maps indicate that DMZ binds to the enzyme in two antiparallel orientations of the long axis of the substrate. One orientation places the principal site of hydroxylation, the 4-methyl group, 4.4 Å from the heme Fe, whereas the alternate conformation positions the second, infrequent site of hydroxylation at &gt;5.9 Å from the heme Fe. Comparison of this structure to that obtained previously for the enzyme indicates that the protein closes around the substrate and prevents open access of water from bulk solvent to the heme Fe. This reflects a ∼1.5 Å movement of the F and G helices relative to helix I. The present structure provides a complete model for the protein from residues 27−488 and defines two new helices F‘ and G‘. The G‘ helix is likely to contribute to interactions of the enzyme with membranes. The relatively large active site, as compared to the volume occupied by the substrate, and the flexibility of the enzyme are likely to underlie the capacity of drug-metabolizing enzymes to metabolize structurally diverse substrates of different sizes.</description><subject>Biochemistry, Molecular Biology</subject><subject>Crystallography, X-Ray</subject><subject>Cytochrome P-450 Enzyme System - chemistry</subject><subject>Cytochrome P450 Family 2</subject><subject>Electrons</subject><subject>Heme - chemistry</subject><subject>Humans</subject><subject>Imidazoles - chemistry</subject><subject>Iron - chemistry</subject><subject>Life Sciences</subject><subject>Models, Molecular</subject><subject>Protein Binding</subject><subject>Protein Conformation</subject><subject>Protein Structure, Tertiary</subject><subject>Steroid 21-Hydroxylase - chemistry</subject><subject>Toxicology</subject><subject>Water - chemistry</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkM9O3DAQh62qqCy0h75A5UsPPQT8J4kTbhBRKNoViKUSN8uxJ8WQxCvbQXDjgtQ-V9-EJyGrRQsHTqP5zTcz0ofQV0p2KGF0t7aECV4y9gFNaMZIkpZl9hFNCCF5wsqcbKKtEK7HNiUi_YQ2KRO5YLSYoL_z6AcdBw_YNVjh-VCH6FUEXLlu0cLdMp6prlOtVT2u7qPTV951gM_SjGBWZVhFzHY4_v-IzyG4dojW9XtPD__w4a010GvAjfN4NrTRjgfffDiwvbH9HzxzBsJntNGoNsCXl7qNfv88vKiOk-np0a9qf5oonqcxMYYVPK2BAMnTWptaN5lpSFPWnJS5FqLIGWTKgKoLoSGlxGhjOBCqxoQZvo1-rO5eqVYuvO2Uv5dOWXm8P5XLbCmtEDy7pa-s9i4ED816gRK5NC_X5kf224pdDHUH5pV8UT0CyQqwIcLdeq78jcwFF5m8OJvLg0s6uzznJ1KM_PcVr3SQ127w_ajlncfPwwKaRA</recordid><startdate>20030603</startdate><enddate>20030603</enddate><creator>Wester, Michael R</creator><creator>Johnson, Eric F</creator><creator>Marques-Soares, Cristina</creator><creator>Dansette, Patrick M</creator><creator>Mansuy, Daniel</creator><creator>Stout, C. David</creator><general>American Chemical Society</general><scope>BSCLL</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>1XC</scope><orcidid>https://orcid.org/0000-0003-0024-0023</orcidid><orcidid>https://orcid.org/0000-0002-3694-3348</orcidid></search><sort><creationdate>20030603</creationdate><title>Structure of a Substrate Complex of Mammalian Cytochrome P450 2C5 at 2.3 Å Resolution:  Evidence for Multiple Substrate Binding Modes</title><author>Wester, Michael R ; Johnson, Eric F ; Marques-Soares, Cristina ; Dansette, Patrick M ; Mansuy, Daniel ; Stout, C. David</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a364t-dd2834be0e064bcdbcf5df0f9b3096c77862e5adeab87ce410dcdd3e01aab82d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Biochemistry, Molecular Biology</topic><topic>Crystallography, X-Ray</topic><topic>Cytochrome P-450 Enzyme System - chemistry</topic><topic>Cytochrome P450 Family 2</topic><topic>Electrons</topic><topic>Heme - chemistry</topic><topic>Humans</topic><topic>Imidazoles - chemistry</topic><topic>Iron - chemistry</topic><topic>Life Sciences</topic><topic>Models, Molecular</topic><topic>Protein Binding</topic><topic>Protein Conformation</topic><topic>Protein Structure, Tertiary</topic><topic>Steroid 21-Hydroxylase - chemistry</topic><topic>Toxicology</topic><topic>Water - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wester, Michael R</creatorcontrib><creatorcontrib>Johnson, Eric F</creatorcontrib><creatorcontrib>Marques-Soares, Cristina</creatorcontrib><creatorcontrib>Dansette, Patrick M</creatorcontrib><creatorcontrib>Mansuy, Daniel</creatorcontrib><creatorcontrib>Stout, C. David</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Hyper Article en Ligne (HAL)</collection><jtitle>Biochemistry (Easton)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wester, Michael R</au><au>Johnson, Eric F</au><au>Marques-Soares, Cristina</au><au>Dansette, Patrick M</au><au>Mansuy, Daniel</au><au>Stout, C. David</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Structure of a Substrate Complex of Mammalian Cytochrome P450 2C5 at 2.3 Å Resolution:  Evidence for Multiple Substrate Binding Modes</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>2003-06-03</date><risdate>2003</risdate><volume>42</volume><issue>21</issue><spage>6370</spage><epage>6379</epage><pages>6370-6379</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>The structure of rabbit microsomal cytochrome P450 2C5/3LVdH complexed with a substrate, 4-methyl-N-methyl-N-(2-phenyl-2H-pyrazol-3-yl)benzenesulfonamide (DMZ), was determined by X-ray crystallography to 2.3 Å resolution. Substrate docking studies and electron density maps indicate that DMZ binds to the enzyme in two antiparallel orientations of the long axis of the substrate. One orientation places the principal site of hydroxylation, the 4-methyl group, 4.4 Å from the heme Fe, whereas the alternate conformation positions the second, infrequent site of hydroxylation at &gt;5.9 Å from the heme Fe. Comparison of this structure to that obtained previously for the enzyme indicates that the protein closes around the substrate and prevents open access of water from bulk solvent to the heme Fe. This reflects a ∼1.5 Å movement of the F and G helices relative to helix I. The present structure provides a complete model for the protein from residues 27−488 and defines two new helices F‘ and G‘. The G‘ helix is likely to contribute to interactions of the enzyme with membranes. The relatively large active site, as compared to the volume occupied by the substrate, and the flexibility of the enzyme are likely to underlie the capacity of drug-metabolizing enzymes to metabolize structurally diverse substrates of different sizes.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>12767218</pmid><doi>10.1021/bi0273922</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0003-0024-0023</orcidid><orcidid>https://orcid.org/0000-0002-3694-3348</orcidid></addata></record>
fulltext fulltext
identifier ISSN: 0006-2960
ispartof Biochemistry (Easton), 2003-06, Vol.42 (21), p.6370-6379
issn 0006-2960
1520-4995
language eng
recordid cdi_hal_primary_oai_HAL_hal_00068735v1
source American Chemical Society; MEDLINE
subjects Biochemistry, Molecular Biology
Crystallography, X-Ray
Cytochrome P-450 Enzyme System - chemistry
Cytochrome P450 Family 2
Electrons
Heme - chemistry
Humans
Imidazoles - chemistry
Iron - chemistry
Life Sciences
Models, Molecular
Protein Binding
Protein Conformation
Protein Structure, Tertiary
Steroid 21-Hydroxylase - chemistry
Toxicology
Water - chemistry
title Structure of a Substrate Complex of Mammalian Cytochrome P450 2C5 at 2.3 Å Resolution:  Evidence for Multiple Substrate Binding Modes
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-03T18%3A43%3A05IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-acs_hal_p&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Structure%20of%20a%20Substrate%20Complex%20of%20Mammalian%20Cytochrome%20P450%202C5%20at%202.3%20%C3%85%20Resolution:%E2%80%89%20Evidence%20for%20Multiple%20Substrate%20Binding%20Modes&rft.jtitle=Biochemistry%20(Easton)&rft.au=Wester,%20Michael%20R&rft.date=2003-06-03&rft.volume=42&rft.issue=21&rft.spage=6370&rft.epage=6379&rft.pages=6370-6379&rft.issn=0006-2960&rft.eissn=1520-4995&rft_id=info:doi/10.1021/bi0273922&rft_dat=%3Cacs_hal_p%3Ea52673040%3C/acs_hal_p%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_id=info:pmid/12767218&rfr_iscdi=true