Immunogold Localisation of the Intermediate Chain within the Protein Complex of Cytoplasmic Dynein
It was our goal to determine the location of the intermediate chain within the complex of cytoplasmic dynein by immunoelectron microscopy. To do so we generated two monoclonal antibodies (m74-1 and m74-2) specific for the intermediate chain. Both antibodies recognised the intermediate chain by sodiu...
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| Veröffentlicht in: | Journal of structural biology 1996-11, Vol.117 (3), p.227-235 |
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| container_title | Journal of structural biology |
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| creator | Steffen, Walter Hodgkinson, Julie L. Wiche, Gerhard |
| description | It was our goal to determine the location of the intermediate chain within the complex of cytoplasmic dynein by immunoelectron microscopy. To do so we generated two monoclonal antibodies (m74-1 and m74-2) specific for the intermediate chain. Both antibodies recognised the intermediate chain by sodium dodecyl sulphate–polyacrylamide gel electrophoresis immunoblot and ELISA assays of native and denatured proteins. When sucrose density gradient-purified cytoplasmic dynein from bovine brain was incubated with the gold-conjugated monoclonal antibodies, m74-1 and m74-2, and examined by negative staining, the gold label was found opposite the globular heads at the base of the V-shaped stalk of the motor complex. The labelling of the intermediate chain is the first mapping of a component within cytoplasmic dynein. The identification of the intermediate chain at the base of the complex supports a possible docking function of the intermediate chain. |
| doi_str_mv | 10.1006/jsbi.1996.0087 |
| format | Article |
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To do so we generated two monoclonal antibodies (m74-1 and m74-2) specific for the intermediate chain. Both antibodies recognised the intermediate chain by sodium dodecyl sulphate–polyacrylamide gel electrophoresis immunoblot and ELISA assays of native and denatured proteins. When sucrose density gradient-purified cytoplasmic dynein from bovine brain was incubated with the gold-conjugated monoclonal antibodies, m74-1 and m74-2, and examined by negative staining, the gold label was found opposite the globular heads at the base of the V-shaped stalk of the motor complex. The labelling of the intermediate chain is the first mapping of a component within cytoplasmic dynein. The identification of the intermediate chain at the base of the complex supports a possible docking function of the intermediate chain.</description><identifier>ISSN: 1047-8477</identifier><identifier>EISSN: 1095-8657</identifier><identifier>DOI: 10.1006/jsbi.1996.0087</identifier><identifier>PMID: 8986653</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Antibodies, Monoclonal ; Brain Chemistry ; Cattle ; Cytoplasm - chemistry ; Cytoplasm - ultrastructure ; Dyneins - chemistry ; Dyneins - immunology ; Dyneins - ultrastructure ; Mice ; Microscopy, Immunoelectron ; Molecular Structure ; Protein Conformation</subject><ispartof>Journal of structural biology, 1996-11, Vol.117 (3), p.227-235</ispartof><rights>1996 Academic Press</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c339t-bd838bcacbaebd3d4710583b71d679b3f30d791aee20dddb210484caf5e8dcc53</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1006/jsbi.1996.0087$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,782,786,3552,27931,27932,46002</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8986653$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Steffen, Walter</creatorcontrib><creatorcontrib>Hodgkinson, Julie L.</creatorcontrib><creatorcontrib>Wiche, Gerhard</creatorcontrib><title>Immunogold Localisation of the Intermediate Chain within the Protein Complex of Cytoplasmic Dynein</title><title>Journal of structural biology</title><addtitle>J Struct Biol</addtitle><description>It was our goal to determine the location of the intermediate chain within the complex of cytoplasmic dynein by immunoelectron microscopy. To do so we generated two monoclonal antibodies (m74-1 and m74-2) specific for the intermediate chain. Both antibodies recognised the intermediate chain by sodium dodecyl sulphate–polyacrylamide gel electrophoresis immunoblot and ELISA assays of native and denatured proteins. When sucrose density gradient-purified cytoplasmic dynein from bovine brain was incubated with the gold-conjugated monoclonal antibodies, m74-1 and m74-2, and examined by negative staining, the gold label was found opposite the globular heads at the base of the V-shaped stalk of the motor complex. The labelling of the intermediate chain is the first mapping of a component within cytoplasmic dynein. The identification of the intermediate chain at the base of the complex supports a possible docking function of the intermediate chain.</description><subject>Animals</subject><subject>Antibodies, Monoclonal</subject><subject>Brain Chemistry</subject><subject>Cattle</subject><subject>Cytoplasm - chemistry</subject><subject>Cytoplasm - ultrastructure</subject><subject>Dyneins - chemistry</subject><subject>Dyneins - immunology</subject><subject>Dyneins - ultrastructure</subject><subject>Mice</subject><subject>Microscopy, Immunoelectron</subject><subject>Molecular Structure</subject><subject>Protein Conformation</subject><issn>1047-8477</issn><issn>1095-8657</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kDtPwzAUhS0E4lFY2ZAysaXYcRI7IwqvSpVggNny44YaJXGxXaD_Hket2JjOlc65RzofQpcEzwnG9c1HUHZOmqaeY8zZAToluKlyXlfscLpLlvOSsRN0FsIHxrgkBTlGx7zhdV3RU6QWw7AZ3bvrTbZ0WvY2yGjdmLkuiyvIFmMEP4CxMkLWrqQds28bV0km98W7COlu3bDu4Wd6arfRrXsZBquzu-2Y3HN01Mk-wMVeZ-jt4f61fcqXz4-L9naZa0qbmCvDKVdaaiVBGWpKRnDFqWLE1KxRtKPYsIZIgAIbY1SRxvFSy64CbrSu6Axd73rX3n1uIEQx2KCh7-UIbhME43VZFQVOwfkuqL0LwUMn1t4O0m8FwWKCKiaoYoIqJqjp4WrfvFGJxV98TzH5fOdDmvdlwYugLYw6YfOgozDO_lf9CxyiiHo</recordid><startdate>199611</startdate><enddate>199611</enddate><creator>Steffen, Walter</creator><creator>Hodgkinson, Julie L.</creator><creator>Wiche, Gerhard</creator><general>Elsevier Inc</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>7X8</scope></search><sort><creationdate>199611</creationdate><title>Immunogold Localisation of the Intermediate Chain within the Protein Complex of Cytoplasmic Dynein</title><author>Steffen, Walter ; Hodgkinson, Julie L. ; Wiche, Gerhard</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c339t-bd838bcacbaebd3d4710583b71d679b3f30d791aee20dddb210484caf5e8dcc53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal</topic><topic>Brain Chemistry</topic><topic>Cattle</topic><topic>Cytoplasm - chemistry</topic><topic>Cytoplasm - ultrastructure</topic><topic>Dyneins - chemistry</topic><topic>Dyneins - immunology</topic><topic>Dyneins - ultrastructure</topic><topic>Mice</topic><topic>Microscopy, Immunoelectron</topic><topic>Molecular Structure</topic><topic>Protein Conformation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Steffen, Walter</creatorcontrib><creatorcontrib>Hodgkinson, Julie L.</creatorcontrib><creatorcontrib>Wiche, Gerhard</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>Journal of structural biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Steffen, Walter</au><au>Hodgkinson, Julie L.</au><au>Wiche, Gerhard</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Immunogold Localisation of the Intermediate Chain within the Protein Complex of Cytoplasmic Dynein</atitle><jtitle>Journal of structural biology</jtitle><addtitle>J Struct Biol</addtitle><date>1996-11</date><risdate>1996</risdate><volume>117</volume><issue>3</issue><spage>227</spage><epage>235</epage><pages>227-235</pages><issn>1047-8477</issn><eissn>1095-8657</eissn><abstract>It was our goal to determine the location of the intermediate chain within the complex of cytoplasmic dynein by immunoelectron microscopy. To do so we generated two monoclonal antibodies (m74-1 and m74-2) specific for the intermediate chain. Both antibodies recognised the intermediate chain by sodium dodecyl sulphate–polyacrylamide gel electrophoresis immunoblot and ELISA assays of native and denatured proteins. When sucrose density gradient-purified cytoplasmic dynein from bovine brain was incubated with the gold-conjugated monoclonal antibodies, m74-1 and m74-2, and examined by negative staining, the gold label was found opposite the globular heads at the base of the V-shaped stalk of the motor complex. The labelling of the intermediate chain is the first mapping of a component within cytoplasmic dynein. The identification of the intermediate chain at the base of the complex supports a possible docking function of the intermediate chain.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>8986653</pmid><doi>10.1006/jsbi.1996.0087</doi><tpages>9</tpages></addata></record> |
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| identifier | ISSN: 1047-8477 |
| ispartof | Journal of structural biology, 1996-11, Vol.117 (3), p.227-235 |
| issn | 1047-8477 1095-8657 |
| language | eng |
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| subjects | Animals Antibodies, Monoclonal Brain Chemistry Cattle Cytoplasm - chemistry Cytoplasm - ultrastructure Dyneins - chemistry Dyneins - immunology Dyneins - ultrastructure Mice Microscopy, Immunoelectron Molecular Structure Protein Conformation |
| title | Immunogold Localisation of the Intermediate Chain within the Protein Complex of Cytoplasmic Dynein |
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