Cross-linking Mass Spectrometry and Mutagenesis Confirm the Functional Importance of Surface Interactions between CYP3A4 and Holo/Apo Cytochrome b5
Identifieur interne : 000517 ( Main/Exploration ); précédent : 000516; suivant : 000518Cross-linking Mass Spectrometry and Mutagenesis Confirm the Functional Importance of Surface Interactions between CYP3A4 and Holo/Apo Cytochrome b5
Auteurs : Chunsheng Zhao [États-Unis] ; Qiuxia Gao [États-Unis] ; Arthur G. Roberts [États-Unis] ; Scott A. Shaffer [États-Unis] ; Catalin E. Doneanu [États-Unis] ; Song Xue [États-Unis] ; David R. Goodlett [États-Unis] ; Sidney D. Nelson [États-Unis] ; William M. Atkins [États-Unis]Source :
- Biochemistry [ 0006-2960 ] ; 2012.
Abstract
Cytochrome
Url:
DOI: 10.1021/bi301069r
PubMed: 23150942
PubMed Central: 3568533
Affiliations:
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Roberts</name><affiliation wicri:level="2"><nlm:aff id="A3">Pharmaceutical and Biomedical Sciences, University of Georgia, Athens, GA 30605</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Géorgie (États-Unis)</region></placeName><wicri:cityArea>Pharmaceutical and Biomedical Sciences, University of Georgia, Athens</wicri:cityArea></affiliation></author><author><name sortKey="Shaffer, Scott A" sort="Shaffer, Scott A" uniqKey="Shaffer S" first="Scott A." last="Shaffer">Scott A. Shaffer</name><affiliation wicri:level="2"><nlm:aff id="A2">Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Shrewsbury, MA 01545</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Massachusetts</region></placeName><wicri:cityArea>Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Shrewsbury</wicri:cityArea></affiliation></author><author><name sortKey="Doneanu, Catalin E" sort="Doneanu, Catalin E" uniqKey="Doneanu C" first="Catalin E." last="Doneanu">Catalin E. 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Atkins</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle, Washington 98195</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Washington (État)</region></placeName><wicri:cityArea>Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle</wicri:cityArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">23150942</idno><idno type="pmc">3568533</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3568533</idno><idno type="RBID">PMC:3568533</idno><idno type="doi">10.1021/bi301069r</idno><date when="2012">2012</date><idno type="wicri:Area/Pmc/Corpus">000423</idno><idno type="wicri:Area/Pmc/Curation">000423</idno><idno type="wicri:Area/Pmc/Checkpoint">000429</idno><idno type="wicri:Area/Ncbi/Merge">000368</idno><idno type="wicri:Area/Ncbi/Curation">000368</idno><idno type="wicri:Area/Ncbi/Checkpoint">000368</idno><idno type="wicri:doubleKey">0006-2960:2012:Zhao C:cross:linking:mass</idno><idno type="wicri:Area/Main/Merge">000518</idno><idno type="wicri:Area/Main/Curation">000517</idno><idno type="wicri:Area/Main/Exploration">000517</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Cross-linking Mass Spectrometry and Mutagenesis Confirm the Functional Importance of Surface Interactions between CYP3A4 and Holo/Apo Cytochrome <italic>b<sub>5</sub></italic></title><author><name sortKey="Zhao, Chunsheng" sort="Zhao, Chunsheng" uniqKey="Zhao C" first="Chunsheng" last="Zhao">Chunsheng Zhao</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle, Washington 98195</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Washington (État)</region></placeName><wicri:cityArea>Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle</wicri:cityArea></affiliation></author><author><name sortKey="Gao, Qiuxia" sort="Gao, Qiuxia" uniqKey="Gao Q" first="Qiuxia" last="Gao">Qiuxia Gao</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle, Washington 98195</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Washington (État)</region></placeName><wicri:cityArea>Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle</wicri:cityArea></affiliation></author><author><name sortKey="Roberts, Arthur G" sort="Roberts, Arthur G" uniqKey="Roberts A" first="Arthur G." last="Roberts">Arthur G. Roberts</name><affiliation wicri:level="2"><nlm:aff id="A3">Pharmaceutical and Biomedical Sciences, University of Georgia, Athens, GA 30605</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Géorgie (États-Unis)</region></placeName><wicri:cityArea>Pharmaceutical and Biomedical Sciences, University of Georgia, Athens</wicri:cityArea></affiliation></author><author><name sortKey="Shaffer, Scott A" sort="Shaffer, Scott A" uniqKey="Shaffer S" first="Scott A." last="Shaffer">Scott A. Shaffer</name><affiliation wicri:level="2"><nlm:aff id="A2">Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Shrewsbury, MA 01545</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Massachusetts</region></placeName><wicri:cityArea>Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Shrewsbury</wicri:cityArea></affiliation></author><author><name sortKey="Doneanu, Catalin E" sort="Doneanu, Catalin E" uniqKey="Doneanu C" first="Catalin E." last="Doneanu">Catalin E. Doneanu</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle, Washington 98195</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Washington (État)</region></placeName><wicri:cityArea>Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle</wicri:cityArea></affiliation></author><author><name sortKey="Xue, Song" sort="Xue, Song" uniqKey="Xue S" first="Song" last="Xue">Song Xue</name><affiliation wicri:level="2"><nlm:aff id="A4">Microsoft Incorporation, 1 Microsoft Way, Redmond, Washington 98052</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Washington (État)</region></placeName><wicri:cityArea>Microsoft Incorporation, 1 Microsoft Way, Redmond</wicri:cityArea></affiliation></author><author><name sortKey="Goodlett, David R" sort="Goodlett, David R" uniqKey="Goodlett D" first="David R." last="Goodlett">David R. Goodlett</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle, Washington 98195</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Washington (État)</region></placeName><wicri:cityArea>Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle</wicri:cityArea></affiliation></author><author><name sortKey="Nelson, Sidney D" sort="Nelson, Sidney D" uniqKey="Nelson S" first="Sidney D." last="Nelson">Sidney D. Nelson</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle, Washington 98195</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Washington (État)</region></placeName><wicri:cityArea>Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle</wicri:cityArea></affiliation></author><author><name sortKey="Atkins, William M" sort="Atkins, William M" uniqKey="Atkins W" first="William M." last="Atkins">William M. Atkins</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle, Washington 98195</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Washington (État)</region></placeName><wicri:cityArea>Department of Medicinal Chemistry, University of Washington, Box 357610, Seattle</wicri:cityArea></affiliation></author></analytic><series><title level="j">Biochemistry</title><idno type="ISSN">0006-2960</idno><idno type="eISSN">1520-4995</idno><imprint><date when="2012">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="P1">Cytochrome <italic>b<sub>5</sub></italic> (cyt <italic>b<sub>5</sub></italic>) is one of the key components in the microsomal cytochrome P450 monooxygenase system. Consensus has not been reached on the underlying mechanism of cyt <italic>b<sub>5</sub></italic> modulation of CYP catalysis. Both cyt <italic>b<sub>5</sub></italic> and apo <italic>b<sub>5</sub></italic>, are reported to stimulate the activity of several P450 isoforms. In the present study, the surface interactions of both holo and apo <italic>b<sub>5</sub></italic> with CYP3A4 were investigated and compared for the first time. Chemical cross-linking coupled with mass spectrometric analysis was used to identify the potential electrostatic interactions between the protein surfaces. Subsequently, the interaction models of holo/apo <italic>b<sub>5</sub></italic> with CYP3A4 were built using the identified interacting sites as constraints. Both cyt <italic>b<sub>5</sub></italic> and apo <italic>b<sub>5</sub></italic> were predicted to bind to the same groove on CYP3A4 with close contacts to the B-B’ loop of CYP3A4, a substrate recognition site (SRS). Mutagenesis studies further confirmed that the interacting sites on CYP3A4 (Lys96, Lys127 and Lys421) are of functional importance. Mutation of these residues reduced or abolished cyt <italic>b<sub>5</sub></italic> binding affinity. The critical role of Arg446 on CYP3A4 in binding to cyt <italic>b<sub>5</sub></italic> and/or cytochrome P450 reductase (CPR) was also discovered. The results indicated that electrostatic interactions on the interface of the two proteins are functionally important. The results indicate that the apo cyt <italic>b</italic><sub>5</sub> can dock with CYP3A4 in a manner analogous to holo cyt <italic>b</italic><sub>5</sub> so electron transfer from cyt <italic>b</italic><sub>5</sub> is not required for its effects.</p></div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Géorgie (États-Unis)</li><li>Massachusetts</li><li>Washington (État)</li></region></list><tree><country name="États-Unis"><region name="Washington (État)"><name sortKey="Zhao, Chunsheng" sort="Zhao, Chunsheng" uniqKey="Zhao C" first="Chunsheng" last="Zhao">Chunsheng Zhao</name></region><name sortKey="Atkins, William M" sort="Atkins, William M" uniqKey="Atkins W" first="William M." last="Atkins">William M. Atkins</name><name sortKey="Doneanu, Catalin E" sort="Doneanu, Catalin E" uniqKey="Doneanu C" first="Catalin E." last="Doneanu">Catalin E. Doneanu</name><name sortKey="Gao, Qiuxia" sort="Gao, Qiuxia" uniqKey="Gao Q" first="Qiuxia" last="Gao">Qiuxia Gao</name><name sortKey="Goodlett, David R" sort="Goodlett, David R" uniqKey="Goodlett D" first="David R." last="Goodlett">David R. Goodlett</name><name sortKey="Nelson, Sidney D" sort="Nelson, Sidney D" uniqKey="Nelson S" first="Sidney D." last="Nelson">Sidney D. Nelson</name><name sortKey="Roberts, Arthur G" sort="Roberts, Arthur G" uniqKey="Roberts A" first="Arthur G." last="Roberts">Arthur G. Roberts</name><name sortKey="Shaffer, Scott A" sort="Shaffer, Scott A" uniqKey="Shaffer S" first="Scott A." last="Shaffer">Scott A. Shaffer</name><name sortKey="Xue, Song" sort="Xue, Song" uniqKey="Xue S" first="Song" last="Xue">Song Xue</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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