Investigation of the binding interactions of progesterone using muteins of the human progesterone receptor ligand binding domain designed on the basis of a three-dimensional protein model
Identifieur interne : 000B36 ( Main/Corpus ); précédent : 000B35; suivant : 000B37Investigation of the binding interactions of progesterone using muteins of the human progesterone receptor ligand binding domain designed on the basis of a three-dimensional protein model
Auteurs : Miriam Letz ; Peter Bringmann ; Mario Mann ; Anke Mueller-Fahrnow ; Dania Reipert ; Peter Scholz ; Jean-Marie Wurtz ; Ursula EgnerSource :
- Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology [ 0167-4838 ] ; 1998.
Abstract
The aim of this study was to investigate the binding interactions of the human progesterone receptor (hPR) with its natural ligand. Therefore, a homology-derived model of the hPR ligand binding domain has been constructed and used to predict residues potentially involved in interactions with progesterone. These residues and the free cysteines have been mutated (in total 13 residues with 15 mutations). All exchanges have been designed to preserve the three-dimensional structure of the protein. With respect to the binding characteristics towards progesterone, the muteins fall into three groups displaying no, reduced, or wildtype-like binding activity.
Url:
DOI: 10.1016/S0167-4838(98)00249-0
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D-13342 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Mueller Fahrnow, Anke" sort="Mueller Fahrnow, Anke" uniqKey="Mueller Fahrnow A" first="Anke" last="Mueller-Fahrnow">Anke Mueller-Fahrnow</name><affiliation><mods:affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Reipert, Dania" sort="Reipert, Dania" uniqKey="Reipert D" first="Dania" last="Reipert">Dania Reipert</name><affiliation><mods:affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Scholz, Peter" sort="Scholz, Peter" uniqKey="Scholz P" first="Peter" last="Scholz">Peter Scholz</name><affiliation><mods:affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Wurtz, Jean Marie" sort="Wurtz, Jean Marie" uniqKey="Wurtz J" first="Jean-Marie" last="Wurtz">Jean-Marie Wurtz</name><affiliation><mods:affiliation>IGBMC, Laboratoire de Biologie Structurale, 1, rue Laurent Fries, BP 163, F-67404 Illkirch, France</mods:affiliation></affiliation></author><author><name sortKey="Egner, Ursula" sort="Egner, Ursula" uniqKey="Egner U" first="Ursula" last="Egner">Ursula Egner</name><affiliation><mods:affiliation>E-mail: ursula.egner@schering.de</mods:affiliation></affiliation><affiliation><mods:affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:E6CCDF1A153759B65F515C74446768BE4C334702</idno><date when="1999" year="1999">1999</date><idno type="doi">10.1016/S0167-4838(98)00249-0</idno><idno type="url">https://api.istex.fr/document/E6CCDF1A153759B65F515C74446768BE4C334702/fulltext/pdf</idno><idno 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Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Mueller Fahrnow, Anke" sort="Mueller Fahrnow, Anke" uniqKey="Mueller Fahrnow A" first="Anke" last="Mueller-Fahrnow">Anke Mueller-Fahrnow</name><affiliation><mods:affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Reipert, Dania" sort="Reipert, Dania" uniqKey="Reipert D" first="Dania" last="Reipert">Dania Reipert</name><affiliation><mods:affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Scholz, Peter" sort="Scholz, Peter" uniqKey="Scholz P" first="Peter" last="Scholz">Peter Scholz</name><affiliation><mods:affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Wurtz, Jean Marie" sort="Wurtz, Jean Marie" uniqKey="Wurtz J" first="Jean-Marie" last="Wurtz">Jean-Marie Wurtz</name><affiliation><mods:affiliation>IGBMC, Laboratoire de Biologie Structurale, 1, rue Laurent Fries, BP 163, F-67404 Illkirch, France</mods:affiliation></affiliation></author><author><name sortKey="Egner, Ursula" sort="Egner, Ursula" uniqKey="Egner U" first="Ursula" last="Egner">Ursula Egner</name><affiliation><mods:affiliation>E-mail: ursula.egner@schering.de</mods:affiliation></affiliation><affiliation><mods:affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology</title><title level="j" type="abbrev">BBAPRO</title><idno type="ISSN">0167-4838</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1998">1998</date><biblScope unit="volume">1429</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="391">391</biblScope><biblScope unit="page" to="400">400</biblScope></imprint><idno type="ISSN">0167-4838</idno></series><idno type="istex">E6CCDF1A153759B65F515C74446768BE4C334702</idno><idno type="DOI">10.1016/S0167-4838(98)00249-0</idno><idno type="PII">S0167-4838(98)00249-0</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0167-4838</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The aim of this study was to investigate the binding interactions of the human progesterone receptor (hPR) with its natural ligand. Therefore, a homology-derived model of the hPR ligand binding domain has been constructed and used to predict residues potentially involved in interactions with progesterone. These residues and the free cysteines have been mutated (in total 13 residues with 15 mutations). All exchanges have been designed to preserve the three-dimensional structure of the protein. With respect to the binding characteristics towards progesterone, the muteins fall into three groups displaying no, reduced, or wildtype-like binding activity.</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>Miriam Letz</name><affiliations><json:string>Research Laboratories of Schering AG, D-13342 Berlin, Germany</json:string></affiliations></json:item><json:item><name>Peter Bringmann</name><affiliations><json:string>Research Laboratories of Schering AG, D-13342 Berlin, Germany</json:string></affiliations></json:item><json:item><name>Mario Mann</name><affiliations><json:string>Research Laboratories of Schering AG, D-13342 Berlin, Germany</json:string></affiliations></json:item><json:item><name>Anke Mueller-Fahrnow</name><affiliations><json:string>Research Laboratories of Schering AG, D-13342 Berlin, Germany</json:string></affiliations></json:item><json:item><name>Dania Reipert</name><affiliations><json:string>Research Laboratories of Schering AG, D-13342 Berlin, Germany</json:string></affiliations></json:item><json:item><name>Peter Scholz</name><affiliations><json:string>Research Laboratories of Schering AG, D-13342 Berlin, Germany</json:string></affiliations></json:item><json:item><name>Jean-Marie Wurtz</name><affiliations><json:string>IGBMC, Laboratoire de Biologie Structurale, 1, rue Laurent Fries, BP 163, F-67404 Illkirch, France</json:string></affiliations></json:item><json:item><name>Ursula Egner</name><affiliations><json:string>E-mail: ursula.egner@schering.de</json:string><json:string>Research Laboratories of Schering AG, D-13342 Berlin, Germany</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>Progesterone receptor</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Ligand binding domain</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Comparative modelling</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Site-directed mutagenesis</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Binding characteristic</value></json:item></subject><language><json:string>eng</json:string></language><abstract>The aim of this study was to investigate the binding interactions of the human progesterone receptor (hPR) with its natural ligand. Therefore, a homology-derived model of the hPR ligand binding domain has been constructed and used to predict residues potentially involved in interactions with progesterone. These residues and the free cysteines have been mutated (in total 13 residues with 15 mutations). All exchanges have been designed to preserve the three-dimensional structure of the protein. With respect to the binding characteristics towards progesterone, the muteins fall into three groups displaying no, reduced, or wildtype-like binding activity.</abstract><qualityIndicators><score>5.822</score><pdfVersion>1.2</pdfVersion><pdfPageSize>588 x 794 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>5</keywordCount><abstractCharCount>657</abstractCharCount><pdfWordCount>4682</pdfWordCount><pdfCharCount>27570</pdfCharCount><pdfPageCount>10</pdfPageCount><abstractWordCount>95</abstractWordCount></qualityIndicators><title>Investigation of the binding interactions of progesterone using muteins of the human progesterone receptor ligand binding domain designed on the basis of a three-dimensional protein model</title><pii><json:string>S0167-4838(98)00249-0</json:string></pii><genre><json:string>research-article</json:string></genre><host><volume>1429</volume><pii><json:string>S0167-4838(00)X0059-3</json:string></pii><pages><last>400</last><first>391</first></pages><issn><json:string>0167-4838</json:string></issn><issue>2</issue><genre></genre><language><json:string>unknown</json:string></language><title>Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular Enzymology</title><publicationDate>1999</publicationDate></host><categories><wos><json:string>BIOCHEMISTRY & MOLECULAR BIOLOGY</json:string><json:string>BIOPHYSICS</json:string></wos></categories><publicationDate>1998</publicationDate><copyrightDate>1999</copyrightDate><doi><json:string>10.1016/S0167-4838(98)00249-0</json:string></doi><id>E6CCDF1A153759B65F515C74446768BE4C334702</id><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/E6CCDF1A153759B65F515C74446768BE4C334702/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/E6CCDF1A153759B65F515C74446768BE4C334702/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/E6CCDF1A153759B65F515C74446768BE4C334702/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a">Investigation of the binding interactions of progesterone using muteins of the human progesterone receptor ligand binding domain designed on the basis of a three-dimensional protein model</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>ELSEVIER</publisher><availability><p>ELSEVIER</p></availability><date>1999</date></publicationStmt><notesStmt><note type="content">Fig. 1: Sequence alignment of progesterone receptor LBDs from various organisms. The sequences of the hERα and hRARγ for which crystal structures have been determined, are also included. Residues identical in the hPR LBDs of different species are coloured green or blue. Similar amino acid residues in all receptors are coloured yellow. The secondary structure information shown below the alignment corresponds to the hRARγ crystal structure. The residues closer than 5 Å to the ligands are indicated by bullets for the hRARγ/all-trans retinoic acid complexes. Species abbreviation are: hs, Homo sapiens; rn, Rattus norvegicus (rat); mm, Mus musculus (mouse); oc, Oryctolagus cuniculus (rabbit); gg, Gallus gallus (chicken); oa, Ovus aries (sheep); cu, Cnemidophorus uniparens (whiptail lizard). The figure was prepared using ALSCRIPT [28].</note><note type="content">Fig. 2: Presentation of the hPR model. (A) Picture showing the overall fold of the hPR LBD with α-helices shown as cylinders. The β-strands are not visible in this orientation. Progesterone is depicted as a CPK model. (B) Positions of the mutated residues in the model of the hPR LBD. Mutated residues with a strong effect on ligand binding are shown in red, those with a moderate effect are coloured cyan and those with no effect in yellow. Ser792, Ser793 and Tyr795 are omitted for clarity. Hydrogen bonds are visualised in cyan.</note><note type="content">Fig. 3: Effects of the directed mutations on the binding activity of the hPR LBD towards progesterone: relative binding affinities. The results of the ligand binding assays with the crude bacterial extracts containing the various muteins were obtained under ligand saturation conditions and are expressed as percentage of the specific binding seen with the wildtype extract. Data represent the mean±S.D. of three independent experiments. As in Fig. 2, the mutations that have a strong, a moderate and no effect on the overall binding activity (considering also KD and half-life values) are shown in red, in cyan and in yellow, respectively.</note><note type="content">Table 1: Effects of the directed mutations on the binding activity of the hPR LBD towards progesterone: KD values and dissociation rate kinetics</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">Investigation of the binding interactions of progesterone using muteins of the human progesterone receptor ligand binding domain designed on the basis of a three-dimensional protein model</title><author><persName><forename type="first">Miriam</forename><surname>Letz</surname></persName><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation></author><author><persName><forename type="first">Peter</forename><surname>Bringmann</surname></persName><note type="biography">Present address: Berlex Biosiences, 15049 San Pablo Avenue, P.O. Box 4099, Richmond, CA 94804-0099, USA.</note><affiliation>Present address: Berlex Biosiences, 15049 San Pablo Avenue, P.O. Box 4099, Richmond, CA 94804-0099, USA.</affiliation><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation></author><author><persName><forename type="first">Mario</forename><surname>Mann</surname></persName><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation></author><author><persName><forename type="first">Anke</forename><surname>Mueller-Fahrnow</surname></persName><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation></author><author><persName><forename type="first">Dania</forename><surname>Reipert</surname></persName><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation></author><author><persName><forename type="first">Peter</forename><surname>Scholz</surname></persName><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation></author><author><persName><forename type="first">Jean-Marie</forename><surname>Wurtz</surname></persName><affiliation>IGBMC, Laboratoire de Biologie Structurale, 1, rue Laurent Fries, BP 163, F-67404 Illkirch, France</affiliation></author><author><persName><forename type="first">Ursula</forename><surname>Egner</surname></persName><email>ursula.egner@schering.de</email><note type="correspondence"><p>Corresponding author. Fax: +49 (30) 4681-6741</p></note><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation></author></analytic><monogr><title level="j">Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology</title><title level="j" type="abbrev">BBAPRO</title><idno type="pISSN">0167-4838</idno><idno type="PII">S0167-4838(00)X0059-3</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1998"></date><biblScope unit="volume">1429</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="391">391</biblScope><biblScope unit="page" to="400">400</biblScope></imprint></monogr><idno type="istex">E6CCDF1A153759B65F515C74446768BE4C334702</idno><idno type="DOI">10.1016/S0167-4838(98)00249-0</idno><idno type="PII">S0167-4838(98)00249-0</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1999</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>The aim of this study was to investigate the binding interactions of the human progesterone receptor (hPR) with its natural ligand. Therefore, a homology-derived model of the hPR ligand binding domain has been constructed and used to predict residues potentially involved in interactions with progesterone. These residues and the free cysteines have been mutated (in total 13 residues with 15 mutations). All exchanges have been designed to preserve the three-dimensional structure of the protein. With respect to the binding characteristics towards progesterone, the muteins fall into three groups displaying no, reduced, or wildtype-like binding activity.</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>Progesterone receptor</term></item><item><term>Ligand binding domain</term></item><item><term>Comparative modelling</term></item><item><term>Site-directed mutagenesis</term></item><item><term>Binding characteristic</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1998-10-22">Registration</change><change when="1998-10-22">Modified</change><change when="1998">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/E6CCDF1A153759B65F515C74446768BE4C334702/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: ce:floats; body; tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"><istex:entity SYSTEM="gr1" NDATA="IMAGE" name="gr1"></istex:entity><istex:entity SYSTEM="gr2" NDATA="IMAGE" name="gr2"></istex:entity><istex:entity SYSTEM="gr3" NDATA="IMAGE" name="gr3"></istex:entity></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>BBAPRO</jid><aid>35810</aid><ce:pii>S0167-4838(98)00249-0</ce:pii><ce:doi>10.1016/S0167-4838(98)00249-0</ce:doi><ce:copyright type="full-transfer" year="1999">Elsevier Science B.V.</ce:copyright></item-info><head><ce:title>Investigation of the binding interactions of progesterone using muteins of the human progesterone receptor ligand binding domain designed on the basis of a three-dimensional protein model</ce:title><ce:author-group><ce:author><ce:given-name>Miriam</ce:given-name><ce:surname>Letz</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Peter</ce:given-name><ce:surname>Bringmann</ce:surname><ce:cross-ref refid="FN1"><ce:sup>1</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Mario</ce:given-name><ce:surname>Mann</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Anke</ce:given-name><ce:surname>Mueller-Fahrnow</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Dania</ce:given-name><ce:surname>Reipert</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Peter</ce:given-name><ce:surname>Scholz</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Jean-Marie</ce:given-name><ce:surname>Wurtz</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Ursula</ce:given-name><ce:surname>Egner</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref><ce:cross-ref refid="COR1">*</ce:cross-ref><ce:e-address>ursula.egner@schering.de</ce:e-address></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Research Laboratories of Schering AG, D-13342 Berlin, Germany</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>IGBMC, Laboratoire de Biologie Structurale, 1, rue Laurent Fries, BP 163, F-67404 Illkirch, France</ce:textfn></ce:affiliation><ce:correspondence id="COR1"><ce:label>*</ce:label><ce:text>Corresponding author. Fax: +49 (30) 4681-6741</ce:text></ce:correspondence><ce:footnote id="FN1"><ce:label>1</ce:label><ce:note-para>Present address: Berlex Biosiences, 15049 San Pablo Avenue, P.O. Box 4099, Richmond, CA 94804-0099, USA.</ce:note-para></ce:footnote></ce:author-group><ce:date-received day="20" month="7" year="1998"></ce:date-received><ce:date-revised day="22" month="10" year="1998"></ce:date-revised><ce:date-accepted day="22" month="10" year="1998"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>The aim of this study was to investigate the binding interactions of the human progesterone receptor (hPR) with its natural ligand. Therefore, a homology-derived model of the hPR ligand binding domain has been constructed and used to predict residues potentially involved in interactions with progesterone. These residues and the free cysteines have been mutated (in total 13 residues with 15 mutations). All exchanges have been designed to preserve the three-dimensional structure of the protein. With respect to the binding characteristics towards progesterone, the muteins fall into three groups displaying no, reduced, or wildtype-like binding activity.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>Progesterone receptor</ce:text></ce:keyword><ce:keyword><ce:text>Ligand binding domain</ce:text></ce:keyword><ce:keyword><ce:text>Comparative modelling</ce:text></ce:keyword><ce:keyword><ce:text>Site-directed mutagenesis</ce:text></ce:keyword><ce:keyword><ce:text>Binding characteristic</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Investigation of the binding interactions of progesterone using muteins of the human progesterone receptor ligand binding domain designed on the basis of a three-dimensional protein model</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Investigation of the binding interactions of progesterone using muteins of the human progesterone receptor ligand binding domain designed on the basis of a three-dimensional protein model</title></titleInfo><name type="personal"><namePart type="given">Miriam</namePart><namePart type="family">Letz</namePart><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Peter</namePart><namePart type="family">Bringmann</namePart><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation><description>Present address: Berlex Biosiences, 15049 San Pablo Avenue, P.O. Box 4099, Richmond, CA 94804-0099, USA.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Mario</namePart><namePart type="family">Mann</namePart><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Anke</namePart><namePart type="family">Mueller-Fahrnow</namePart><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Dania</namePart><namePart type="family">Reipert</namePart><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Peter</namePart><namePart type="family">Scholz</namePart><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Jean-Marie</namePart><namePart type="family">Wurtz</namePart><affiliation>IGBMC, Laboratoire de Biologie Structurale, 1, rue Laurent Fries, BP 163, F-67404 Illkirch, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Ursula</namePart><namePart type="family">Egner</namePart><affiliation>E-mail: ursula.egner@schering.de</affiliation><affiliation>Research Laboratories of Schering AG, D-13342 Berlin, Germany</affiliation><description>Corresponding author. Fax: +49 (30) 4681-6741</description><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article"></genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1998</dateIssued><dateValid encoding="w3cdtf">1998-10-22</dateValid><dateModified encoding="w3cdtf">1998-10-22</dateModified><copyrightDate encoding="w3cdtf">1999</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">The aim of this study was to investigate the binding interactions of the human progesterone receptor (hPR) with its natural ligand. Therefore, a homology-derived model of the hPR ligand binding domain has been constructed and used to predict residues potentially involved in interactions with progesterone. These residues and the free cysteines have been mutated (in total 13 residues with 15 mutations). All exchanges have been designed to preserve the three-dimensional structure of the protein. With respect to the binding characteristics towards progesterone, the muteins fall into three groups displaying no, reduced, or wildtype-like binding activity.</abstract><note type="content">Fig. 1: Sequence alignment of progesterone receptor LBDs from various organisms. The sequences of the hERα and hRARγ for which crystal structures have been determined, are also included. Residues identical in the hPR LBDs of different species are coloured green or blue. Similar amino acid residues in all receptors are coloured yellow. The secondary structure information shown below the alignment corresponds to the hRARγ crystal structure. The residues closer than 5 Å to the ligands are indicated by bullets for the hRARγ/all-trans retinoic acid complexes. Species abbreviation are: hs, Homo sapiens; rn, Rattus norvegicus (rat); mm, Mus musculus (mouse); oc, Oryctolagus cuniculus (rabbit); gg, Gallus gallus (chicken); oa, Ovus aries (sheep); cu, Cnemidophorus uniparens (whiptail lizard). The figure was prepared using ALSCRIPT [28].</note><note type="content">Fig. 2: Presentation of the hPR model. (A) Picture showing the overall fold of the hPR LBD with α-helices shown as cylinders. The β-strands are not visible in this orientation. Progesterone is depicted as a CPK model. (B) Positions of the mutated residues in the model of the hPR LBD. Mutated residues with a strong effect on ligand binding are shown in red, those with a moderate effect are coloured cyan and those with no effect in yellow. Ser792, Ser793 and Tyr795 are omitted for clarity. Hydrogen bonds are visualised in cyan.</note><note type="content">Fig. 3: Effects of the directed mutations on the binding activity of the hPR LBD towards progesterone: relative binding affinities. The results of the ligand binding assays with the crude bacterial extracts containing the various muteins were obtained under ligand saturation conditions and are expressed as percentage of the specific binding seen with the wildtype extract. Data represent the mean±S.D. of three independent experiments. As in Fig. 2, the mutations that have a strong, a moderate and no effect on the overall binding activity (considering also KD and half-life values) are shown in red, in cyan and in yellow, respectively.</note><note type="content">Table 1: Effects of the directed mutations on the binding activity of the hPR LBD towards progesterone: KD values and dissociation rate kinetics</note><subject><genre>Keywords</genre><topic>Progesterone receptor</topic><topic>Ligand binding domain</topic><topic>Comparative modelling</topic><topic>Site-directed mutagenesis</topic><topic>Binding characteristic</topic></subject><relatedItem type="host"><titleInfo><title>Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology</title></titleInfo><titleInfo type="abbreviated"><title>BBAPRO</title></titleInfo><genre type="Journal">journal</genre><originInfo><dateIssued encoding="w3cdtf">19990111</dateIssued></originInfo><identifier type="ISSN">0167-4838</identifier><identifier type="PII">S0167-4838(00)X0059-3</identifier><part><date>19990111</date><detail type="volume"><number>1429</number><caption>vol.</caption></detail><detail type="issue"><number>2</number><caption>no.</caption></detail><extent unit="issue pages"><start>293</start><end>528</end></extent><extent unit="pages"><start>391</start><end>400</end></extent></part></relatedItem><identifier type="istex">E6CCDF1A153759B65F515C74446768BE4C334702</identifier><identifier type="DOI">10.1016/S0167-4838(98)00249-0</identifier><identifier type="PII">S0167-4838(98)00249-0</identifier><accessCondition type="use and reproduction" contentType="">© 1999Elsevier Science B.V.</accessCondition><recordInfo><recordContentSource>ELSEVIER</recordContentSource><recordOrigin>Elsevier Science B.V., ©1999</recordOrigin></recordInfo></mods></metadata><enrichments><istex:catWosTEI uri="https://api.istex.fr/document/E6CCDF1A153759B65F515C74446768BE4C334702/enrichments/catWos"><teiHeader><profileDesc><textClass><classCode scheme="WOS">BIOCHEMISTRY & MOLECULAR BIOLOGY</classCode><classCode scheme="WOS">BIOPHYSICS</classCode></textClass></profileDesc></teiHeader></istex:catWosTEI></enrichments><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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