Structure determination of the seven-helix transmembrane receptor sensory rhodopsin II by solution NMR spectroscopy
Membrane proteins pose a huge challenge for structural analysis, but a new study reports the first NMR structure determination of a detergent-solubilized seven-helical transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II. This case study may open the doors to similar solution NM...
Gespeichert in:
| Veröffentlicht in: | Nature Structural & Molecular Biology 2010-06, Vol.17 (6), p.768-774 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 774 |
|---|---|
| container_issue | 6 |
| container_start_page | 768 |
| container_title | Nature Structural & Molecular Biology |
| container_volume | 17 |
| creator | Nietlispach, Daniel Mott, Helen R Gautier, Antoine Bostock, Mark J Kirkpatrick, John P |
| description | Membrane proteins pose a huge challenge for structural analysis, but a new study reports the first NMR structure determination of a detergent-solubilized seven-helical transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II. This case study may open the doors to similar solution NMR structures for other 7TM proteins.
Seven-helix membrane proteins represent a challenge for structural biology. Here we report the first NMR structure determination of a detergent-solubilized seven-helix transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II (pSRII) from
Natronomonas pharaonis
, as a proof of principle. The overall quality of the structure ensemble is good (backbone r.m.s. deviation of 0.48 Å) and agrees well with previously determined X-ray structures. Furthermore, measurements in more native-like small phospholipid bicelles indicate that the protein structure is the same as in detergent micelles, suggesting that environment-specific effects are minimal when using mild detergents. We use our case study as a platform to discuss the feasibility of similar solution NMR studies for other 7TM proteins, including members of the family of G protein–coupled receptors. |
| doi_str_mv | 10.1038/nsmb.1807 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_proqu</sourceid><recordid>TN_cdi_proquest_miscellaneous_746011437</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A228716190</galeid><sourcerecordid>A228716190</sourcerecordid><originalsourceid>FETCH-LOGICAL-c604t-6822d210a4aa47d90d5352d72fdca293e77b6e02827debf26569240ad432d8633</originalsourceid><addsrcrecordid>eNqFkltrFDEUxwdR7EUf_AIa9EEUZs11knksxctCVWj1OWSSM7tTZiZjkpHut2_WrVsqC5KHE5Lf-Z8L_6J4QfCCYKY-jHFoFkRh-ag4JoKLsq6VeLy_1-yoOInxGmMqhGRPiyOKBaFE4OMiXqUw2zQHQA4ShKEbTer8iHyL0hpQhN8wlmvouxuUgsmVYGhyBBTAwpR8yMgYfdigsPbOT7Eb0XKJmg2Kvp__SH37eoniBDYFH62fNs-KJ63pIzy_i6fFz08ff5x_KS--f16en12UtsI8lZWi1FGCDTeGS1djJ5igTtLWWUNrBlI2FWCqqHTQtLQSVU05No4z6lTF2Gnxdqc7Bf9rhpj00EULfZ_b93PUkleYEM7k_0nGMqcqkcnX_5DXfg5jHkMrrnK3XG7l3uyglelBd2Pr8-rsVlKfUaokqUiNM7U4QOXjYOisH6Ht8vuDhHcPEjKT4CatzByjXl5dHmRtXnoM0OopdIMJG02w3ppGb02jt6bJ7Mu7meZmALcn_7okA-93QMxf4wrC_dCH1F7t4GykbKu92j1xC3hC1N0</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>848210477</pqid></control><display><type>article</type><title>Structure determination of the seven-helix transmembrane receptor sensory rhodopsin II by solution NMR spectroscopy</title><source>MEDLINE</source><source>SpringerLink Journals</source><source>Nature Journals Online</source><creator>Nietlispach, Daniel ; Mott, Helen R ; Gautier, Antoine ; Bostock, Mark J ; Kirkpatrick, John P</creator><creatorcontrib>Nietlispach, Daniel ; Mott, Helen R ; Gautier, Antoine ; Bostock, Mark J ; Kirkpatrick, John P</creatorcontrib><description>Membrane proteins pose a huge challenge for structural analysis, but a new study reports the first NMR structure determination of a detergent-solubilized seven-helical transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II. This case study may open the doors to similar solution NMR structures for other 7TM proteins.
Seven-helix membrane proteins represent a challenge for structural biology. Here we report the first NMR structure determination of a detergent-solubilized seven-helix transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II (pSRII) from
Natronomonas pharaonis
, as a proof of principle. The overall quality of the structure ensemble is good (backbone r.m.s. deviation of 0.48 Å) and agrees well with previously determined X-ray structures. Furthermore, measurements in more native-like small phospholipid bicelles indicate that the protein structure is the same as in detergent micelles, suggesting that environment-specific effects are minimal when using mild detergents. We use our case study as a platform to discuss the feasibility of similar solution NMR studies for other 7TM proteins, including members of the family of G protein–coupled receptors.</description><identifier>ISSN: 1545-9993</identifier><identifier>ISSN: 1545-9985</identifier><identifier>EISSN: 1545-9985</identifier><identifier>DOI: 10.1038/nsmb.1807</identifier><identifier>PMID: 20512150</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>631/1647/2258 ; 631/45/535/878 ; Biochemistry ; Biological Microscopy ; Biomedical and Life Sciences ; Case studies ; Cell receptors ; Detergents ; Halobacteriaceae - chemistry ; Halorhodopsins - chemistry ; Hydrophobic and Hydrophilic Interactions ; Life Sciences ; Membrane Biology ; Membrane proteins ; Membranes ; Micelles ; Models, Molecular ; Molecular biology ; Molecular structure ; NMR ; Nuclear magnetic resonance ; Nuclear magnetic resonance spectroscopy ; Nuclear Magnetic Resonance, Biomolecular ; Physiological aspects ; Protein Stability ; Protein Structure ; Protein Structure, Secondary ; Proteins ; Receptors, G-Protein-Coupled - chemistry ; Rhodopsin ; Sensory Rhodopsins - chemistry ; Solubility ; Structure ; technical-report ; Thermodynamics</subject><ispartof>Nature Structural & Molecular Biology, 2010-06, Vol.17 (6), p.768-774</ispartof><rights>Springer Nature America, Inc. 2010</rights><rights>COPYRIGHT 2010 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Jun 2010</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c604t-6822d210a4aa47d90d5352d72fdca293e77b6e02827debf26569240ad432d8633</citedby><cites>FETCH-LOGICAL-c604t-6822d210a4aa47d90d5352d72fdca293e77b6e02827debf26569240ad432d8633</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nsmb.1807$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nsmb.1807$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,778,782,27907,27908,41471,42540,51302</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20512150$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nietlispach, Daniel</creatorcontrib><creatorcontrib>Mott, Helen R</creatorcontrib><creatorcontrib>Gautier, Antoine</creatorcontrib><creatorcontrib>Bostock, Mark J</creatorcontrib><creatorcontrib>Kirkpatrick, John P</creatorcontrib><title>Structure determination of the seven-helix transmembrane receptor sensory rhodopsin II by solution NMR spectroscopy</title><title>Nature Structural & Molecular Biology</title><addtitle>Nat Struct Mol Biol</addtitle><addtitle>Nat Struct Mol Biol</addtitle><description>Membrane proteins pose a huge challenge for structural analysis, but a new study reports the first NMR structure determination of a detergent-solubilized seven-helical transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II. This case study may open the doors to similar solution NMR structures for other 7TM proteins.
Seven-helix membrane proteins represent a challenge for structural biology. Here we report the first NMR structure determination of a detergent-solubilized seven-helix transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II (pSRII) from
Natronomonas pharaonis
, as a proof of principle. The overall quality of the structure ensemble is good (backbone r.m.s. deviation of 0.48 Å) and agrees well with previously determined X-ray structures. Furthermore, measurements in more native-like small phospholipid bicelles indicate that the protein structure is the same as in detergent micelles, suggesting that environment-specific effects are minimal when using mild detergents. We use our case study as a platform to discuss the feasibility of similar solution NMR studies for other 7TM proteins, including members of the family of G protein–coupled receptors.</description><subject>631/1647/2258</subject><subject>631/45/535/878</subject><subject>Biochemistry</subject><subject>Biological Microscopy</subject><subject>Biomedical and Life Sciences</subject><subject>Case studies</subject><subject>Cell receptors</subject><subject>Detergents</subject><subject>Halobacteriaceae - chemistry</subject><subject>Halorhodopsins - chemistry</subject><subject>Hydrophobic and Hydrophilic Interactions</subject><subject>Life Sciences</subject><subject>Membrane Biology</subject><subject>Membrane proteins</subject><subject>Membranes</subject><subject>Micelles</subject><subject>Models, Molecular</subject><subject>Molecular biology</subject><subject>Molecular structure</subject><subject>NMR</subject><subject>Nuclear magnetic resonance</subject><subject>Nuclear magnetic resonance spectroscopy</subject><subject>Nuclear Magnetic Resonance, Biomolecular</subject><subject>Physiological aspects</subject><subject>Protein Stability</subject><subject>Protein Structure</subject><subject>Protein Structure, Secondary</subject><subject>Proteins</subject><subject>Receptors, G-Protein-Coupled - chemistry</subject><subject>Rhodopsin</subject><subject>Sensory Rhodopsins - chemistry</subject><subject>Solubility</subject><subject>Structure</subject><subject>technical-report</subject><subject>Thermodynamics</subject><issn>1545-9993</issn><issn>1545-9985</issn><issn>1545-9985</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqFkltrFDEUxwdR7EUf_AIa9EEUZs11knksxctCVWj1OWSSM7tTZiZjkpHut2_WrVsqC5KHE5Lf-Z8L_6J4QfCCYKY-jHFoFkRh-ag4JoKLsq6VeLy_1-yoOInxGmMqhGRPiyOKBaFE4OMiXqUw2zQHQA4ShKEbTer8iHyL0hpQhN8wlmvouxuUgsmVYGhyBBTAwpR8yMgYfdigsPbOT7Eb0XKJmg2Kvp__SH37eoniBDYFH62fNs-KJ63pIzy_i6fFz08ff5x_KS--f16en12UtsI8lZWi1FGCDTeGS1djJ5igTtLWWUNrBlI2FWCqqHTQtLQSVU05No4z6lTF2Gnxdqc7Bf9rhpj00EULfZ_b93PUkleYEM7k_0nGMqcqkcnX_5DXfg5jHkMrrnK3XG7l3uyglelBd2Pr8-rsVlKfUaokqUiNM7U4QOXjYOisH6Ht8vuDhHcPEjKT4CatzByjXl5dHmRtXnoM0OopdIMJG02w3ppGb02jt6bJ7Mu7meZmALcn_7okA-93QMxf4wrC_dCH1F7t4GykbKu92j1xC3hC1N0</recordid><startdate>20100601</startdate><enddate>20100601</enddate><creator>Nietlispach, Daniel</creator><creator>Mott, Helen R</creator><creator>Gautier, Antoine</creator><creator>Bostock, Mark J</creator><creator>Kirkpatrick, John P</creator><general>Nature Publishing Group US</general><general>Nature Publishing Group</general><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>ISR</scope><scope>3V.</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>P64</scope><scope>PADUT</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20100601</creationdate><title>Structure determination of the seven-helix transmembrane receptor sensory rhodopsin II by solution NMR spectroscopy</title><author>Nietlispach, Daniel ; Mott, Helen R ; Gautier, Antoine ; Bostock, Mark J ; Kirkpatrick, John P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c604t-6822d210a4aa47d90d5352d72fdca293e77b6e02827debf26569240ad432d8633</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>631/1647/2258</topic><topic>631/45/535/878</topic><topic>Biochemistry</topic><topic>Biological Microscopy</topic><topic>Biomedical and Life Sciences</topic><topic>Case studies</topic><topic>Cell receptors</topic><topic>Detergents</topic><topic>Halobacteriaceae - chemistry</topic><topic>Halorhodopsins - chemistry</topic><topic>Hydrophobic and Hydrophilic Interactions</topic><topic>Life Sciences</topic><topic>Membrane Biology</topic><topic>Membrane proteins</topic><topic>Membranes</topic><topic>Micelles</topic><topic>Models, Molecular</topic><topic>Molecular biology</topic><topic>Molecular structure</topic><topic>NMR</topic><topic>Nuclear magnetic resonance</topic><topic>Nuclear magnetic resonance spectroscopy</topic><topic>Nuclear Magnetic Resonance, Biomolecular</topic><topic>Physiological aspects</topic><topic>Protein Stability</topic><topic>Protein Structure</topic><topic>Protein Structure, Secondary</topic><topic>Proteins</topic><topic>Receptors, G-Protein-Coupled - chemistry</topic><topic>Rhodopsin</topic><topic>Sensory Rhodopsins - chemistry</topic><topic>Solubility</topic><topic>Structure</topic><topic>technical-report</topic><topic>Thermodynamics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nietlispach, Daniel</creatorcontrib><creatorcontrib>Mott, Helen R</creatorcontrib><creatorcontrib>Gautier, Antoine</creatorcontrib><creatorcontrib>Bostock, Mark J</creatorcontrib><creatorcontrib>Kirkpatrick, John P</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Research Library (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Research Library Prep</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Research Library</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Research Library (Corporate)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Research Library China</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Nature Structural & Molecular Biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nietlispach, Daniel</au><au>Mott, Helen R</au><au>Gautier, Antoine</au><au>Bostock, Mark J</au><au>Kirkpatrick, John P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Structure determination of the seven-helix transmembrane receptor sensory rhodopsin II by solution NMR spectroscopy</atitle><jtitle>Nature Structural & Molecular Biology</jtitle><stitle>Nat Struct Mol Biol</stitle><addtitle>Nat Struct Mol Biol</addtitle><date>2010-06-01</date><risdate>2010</risdate><volume>17</volume><issue>6</issue><spage>768</spage><epage>774</epage><pages>768-774</pages><issn>1545-9993</issn><issn>1545-9985</issn><eissn>1545-9985</eissn><abstract>Membrane proteins pose a huge challenge for structural analysis, but a new study reports the first NMR structure determination of a detergent-solubilized seven-helical transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II. This case study may open the doors to similar solution NMR structures for other 7TM proteins.
Seven-helix membrane proteins represent a challenge for structural biology. Here we report the first NMR structure determination of a detergent-solubilized seven-helix transmembrane (7TM) protein, the phototaxis receptor sensory rhodopsin II (pSRII) from
Natronomonas pharaonis
, as a proof of principle. The overall quality of the structure ensemble is good (backbone r.m.s. deviation of 0.48 Å) and agrees well with previously determined X-ray structures. Furthermore, measurements in more native-like small phospholipid bicelles indicate that the protein structure is the same as in detergent micelles, suggesting that environment-specific effects are minimal when using mild detergents. We use our case study as a platform to discuss the feasibility of similar solution NMR studies for other 7TM proteins, including members of the family of G protein–coupled receptors.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>20512150</pmid><doi>10.1038/nsmb.1807</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1545-9993 |
| ispartof | Nature Structural & Molecular Biology, 2010-06, Vol.17 (6), p.768-774 |
| issn | 1545-9993 1545-9985 1545-9985 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_746011437 |
| source | MEDLINE; SpringerLink Journals; Nature Journals Online |
| subjects | 631/1647/2258 631/45/535/878 Biochemistry Biological Microscopy Biomedical and Life Sciences Case studies Cell receptors Detergents Halobacteriaceae - chemistry Halorhodopsins - chemistry Hydrophobic and Hydrophilic Interactions Life Sciences Membrane Biology Membrane proteins Membranes Micelles Models, Molecular Molecular biology Molecular structure NMR Nuclear magnetic resonance Nuclear magnetic resonance spectroscopy Nuclear Magnetic Resonance, Biomolecular Physiological aspects Protein Stability Protein Structure Protein Structure, Secondary Proteins Receptors, G-Protein-Coupled - chemistry Rhodopsin Sensory Rhodopsins - chemistry Solubility Structure technical-report Thermodynamics |
| title | Structure determination of the seven-helix transmembrane receptor sensory rhodopsin II by solution NMR spectroscopy |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-17T04%3A32%3A35IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_proqu&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Structure%20determination%20of%20the%20seven-helix%20transmembrane%20receptor%20sensory%20rhodopsin%20II%20by%20solution%20NMR%20spectroscopy&rft.jtitle=Nature%20Structural%20&%20Molecular%20Biology&rft.au=Nietlispach,%20Daniel&rft.date=2010-06-01&rft.volume=17&rft.issue=6&rft.spage=768&rft.epage=774&rft.pages=768-774&rft.issn=1545-9993&rft.eissn=1545-9985&rft_id=info:doi/10.1038/nsmb.1807&rft_dat=%3Cgale_proqu%3EA228716190%3C/gale_proqu%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=848210477&rft_id=info:pmid/20512150&rft_galeid=A228716190&rfr_iscdi=true |