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The cytoskeletal protein talin contains at least two distinct vinculin binding domains
We have mapped the vinculin-binding sites in the cytoskeletal protein talin as well as those sequences which target the talin molecule to focal contacts. Using a series of overlapping talin-fusion proteins expressed in E. coli and (125)I-vinculin in both gel-overlay and microtitre well binding assay...
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| Published in: | The Journal of cell biology 1993-07, Vol.122 (2), p.337-347 |
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| Main Authors: | , , , , , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c4597-e34e9645f955acd5a1595d1831eeb3158c86fd4f079801d10e8a774e7c82bcb43 |
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| container_end_page | 347 |
| container_issue | 2 |
| container_start_page | 337 |
| container_title | The Journal of cell biology |
| container_volume | 122 |
| creator | Gilmore, A.P Wood, C Ohanian, V Jackson, P Patel, B Rees, D.J.G Hynes, R.O Critchley, D.R |
| description | We have mapped the vinculin-binding sites in the cytoskeletal protein talin as well as those sequences which target the talin molecule to focal contacts. Using a series of overlapping talin-fusion proteins expressed in E. coli and (125)I-vinculin in both gel-overlay and microtitre well binding assays, we present evidence for three separable binding sites for vinculin. All three are in the tail segment of talin (residues 434-2541) and are recognized by the same fragment of vinculin (residues 1-258). Two sites are adjacent to each other and span residues 498-950, and the third site is more than 700 residues distant in the primary sequence. Scatchard analysis of (125)I-vinculin binding to talin also indicates three sites, each with a similar affinity (Kd = 2-6 x 10-(7) M). We also detect a substoichiometric interaction of higher affinity (Kd = 3 x 10(-8) M) which remains unexplained. By expressing regions of the chicken talin molecule in heterologous cells, we have shown that the sequences required to target talin to focal contacts overlap those which bind vinculin |
| doi_str_mv | 10.1083/jcb.122.2.337 |
| format | article |
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Using a series of overlapping talin-fusion proteins expressed in E. coli and (125)I-vinculin in both gel-overlay and microtitre well binding assays, we present evidence for three separable binding sites for vinculin. All three are in the tail segment of talin (residues 434-2541) and are recognized by the same fragment of vinculin (residues 1-258). Two sites are adjacent to each other and span residues 498-950, and the third site is more than 700 residues distant in the primary sequence. Scatchard analysis of (125)I-vinculin binding to talin also indicates three sites, each with a similar affinity (Kd = 2-6 x 10-(7) M). We also detect a substoichiometric interaction of higher affinity (Kd = 3 x 10(-8) M) which remains unexplained. By expressing regions of the chicken talin molecule in heterologous cells, we have shown that the sequences required to target talin to focal contacts overlap those which bind vinculin</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.122.2.337</identifier><identifier>PMID: 8320257</identifier><identifier>CODEN: JCLBA3</identifier><language>eng</language><publisher>New York, NY: Rockefeller University Press</publisher><subject>3T3 Cells ; Animals ; Binding Sites ; Biological and medical sciences ; Cell Line ; Cell lines ; Cell structures and functions ; Cells ; Cellular biology ; Chickens ; Complementary DNA ; COS cells ; Cytoskeleton, cytoplasm. Intracellular movements ; ESTRUCTURA CELULAR ; Focal adhesions ; Fundamental and applied biological sciences. Psychology ; Integrins ; Mice ; Molecular and cellular biology ; Molecules ; NIH 3T3 cells ; POLLO ; POULET ; Poultry ; PROTEINAS ; PROTEINE ; Proteins ; Recombinant Fusion Proteins - metabolism ; STRUCTURE CELLULAIRE ; Talin - chemistry ; Talin - genetics ; Talin - metabolism ; Transfection ; Vinculin - metabolism</subject><ispartof>The Journal of cell biology, 1993-07, Vol.122 (2), p.337-347</ispartof><rights>Copyright 1993 The Rockefeller University Press</rights><rights>1993 INIST-CNRS</rights><rights>Copyright Rockefeller University Press Jul 1993</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4597-e34e9645f955acd5a1595d1831eeb3158c86fd4f079801d10e8a774e7c82bcb43</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/1615848$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/1615848$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,778,782,883,27907,27908,58221,58454</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4885537$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8320257$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gilmore, A.P</creatorcontrib><creatorcontrib>Wood, C</creatorcontrib><creatorcontrib>Ohanian, V</creatorcontrib><creatorcontrib>Jackson, P</creatorcontrib><creatorcontrib>Patel, B</creatorcontrib><creatorcontrib>Rees, D.J.G</creatorcontrib><creatorcontrib>Hynes, R.O</creatorcontrib><creatorcontrib>Critchley, D.R</creatorcontrib><title>The cytoskeletal protein talin contains at least two distinct vinculin binding domains</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>We have mapped the vinculin-binding sites in the cytoskeletal protein talin as well as those sequences which target the talin molecule to focal contacts. Using a series of overlapping talin-fusion proteins expressed in E. coli and (125)I-vinculin in both gel-overlay and microtitre well binding assays, we present evidence for three separable binding sites for vinculin. All three are in the tail segment of talin (residues 434-2541) and are recognized by the same fragment of vinculin (residues 1-258). Two sites are adjacent to each other and span residues 498-950, and the third site is more than 700 residues distant in the primary sequence. Scatchard analysis of (125)I-vinculin binding to talin also indicates three sites, each with a similar affinity (Kd = 2-6 x 10-(7) M). We also detect a substoichiometric interaction of higher affinity (Kd = 3 x 10(-8) M) which remains unexplained. By expressing regions of the chicken talin molecule in heterologous cells, we have shown that the sequences required to target talin to focal contacts overlap those which bind vinculin</description><subject>3T3 Cells</subject><subject>Animals</subject><subject>Binding Sites</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>Cell lines</subject><subject>Cell structures and functions</subject><subject>Cells</subject><subject>Cellular biology</subject><subject>Chickens</subject><subject>Complementary DNA</subject><subject>COS cells</subject><subject>Cytoskeleton, cytoplasm. Intracellular movements</subject><subject>ESTRUCTURA CELULAR</subject><subject>Focal adhesions</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Integrins</subject><subject>Mice</subject><subject>Molecular and cellular biology</subject><subject>Molecules</subject><subject>NIH 3T3 cells</subject><subject>POLLO</subject><subject>POULET</subject><subject>Poultry</subject><subject>PROTEINAS</subject><subject>PROTEINE</subject><subject>Proteins</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>STRUCTURE CELLULAIRE</subject><subject>Talin - chemistry</subject><subject>Talin - genetics</subject><subject>Talin - metabolism</subject><subject>Transfection</subject><subject>Vinculin - metabolism</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><recordid>eNpdkctvEzEQxi0EKqFw5IJAWiHU2wY_1_YFqap4SZU40HK1vN7Z1GFjB9vbqv99HRKFx2XG0vfT55n5EHpJ8JJgxd6vXb8klC7pkjH5CC2I4LhVhOPHaIExJa0WVDxFz3JeY4y55OwEnShGMRVygX5c3UDj7kvMP2GCYqdmm2IBH5r6rtXFUKwPubGlmcDm0pS72Aw-Fx9caW5rnXdc78Pgw6oZ4maHP0dPRjtleHHop-j608eriy_t5bfPXy_OL1vHhZYtMA6642LUQlg3CEuEFgNRjAD0jAjlVDcOfMRSK0wGgkFZKTlIp2jves5O0Ye973buNzA4CCXZyWyT39h0b6L15l8l-BuzireGEqI7pqrB2cEgxV8z5GI2PjuYJhsgztlIoQgVnazg2__AdZxTqMtVL4k1r6euULuHXIo5JxiPkxBsdmmZmpapaRlqalqVf_P3-Ef6EE_V3x10m52dxmSD8_mIcaWE-G3zeo-tc4npz59dvSDf7fhqL482GrtK1eH6u-a007hjDwZCr64</recordid><startdate>199307</startdate><enddate>199307</enddate><creator>Gilmore, A.P</creator><creator>Wood, C</creator><creator>Ohanian, V</creator><creator>Jackson, P</creator><creator>Patel, B</creator><creator>Rees, D.J.G</creator><creator>Hynes, R.O</creator><creator>Critchley, D.R</creator><general>Rockefeller University Press</general><general>The Rockefeller University Press</general><scope>FBQ</scope><scope>IQODW</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>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>199307</creationdate><title>The cytoskeletal protein talin contains at least two distinct vinculin binding domains</title><author>Gilmore, A.P ; Wood, C ; Ohanian, V ; Jackson, P ; Patel, B ; Rees, D.J.G ; Hynes, R.O ; Critchley, D.R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4597-e34e9645f955acd5a1595d1831eeb3158c86fd4f079801d10e8a774e7c82bcb43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>3T3 Cells</topic><topic>Animals</topic><topic>Binding Sites</topic><topic>Biological and medical sciences</topic><topic>Cell Line</topic><topic>Cell lines</topic><topic>Cell structures and functions</topic><topic>Cells</topic><topic>Cellular biology</topic><topic>Chickens</topic><topic>Complementary DNA</topic><topic>COS cells</topic><topic>Cytoskeleton, cytoplasm. Intracellular movements</topic><topic>ESTRUCTURA CELULAR</topic><topic>Focal adhesions</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Integrins</topic><topic>Mice</topic><topic>Molecular and cellular biology</topic><topic>Molecules</topic><topic>NIH 3T3 cells</topic><topic>POLLO</topic><topic>POULET</topic><topic>Poultry</topic><topic>PROTEINAS</topic><topic>PROTEINE</topic><topic>Proteins</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>STRUCTURE CELLULAIRE</topic><topic>Talin - chemistry</topic><topic>Talin - genetics</topic><topic>Talin - metabolism</topic><topic>Transfection</topic><topic>Vinculin - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gilmore, A.P</creatorcontrib><creatorcontrib>Wood, C</creatorcontrib><creatorcontrib>Ohanian, V</creatorcontrib><creatorcontrib>Jackson, P</creatorcontrib><creatorcontrib>Patel, B</creatorcontrib><creatorcontrib>Rees, D.J.G</creatorcontrib><creatorcontrib>Hynes, R.O</creatorcontrib><creatorcontrib>Critchley, D.R</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</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>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gilmore, A.P</au><au>Wood, C</au><au>Ohanian, V</au><au>Jackson, P</au><au>Patel, B</au><au>Rees, D.J.G</au><au>Hynes, R.O</au><au>Critchley, D.R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The cytoskeletal protein talin contains at least two distinct vinculin binding domains</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>1993-07</date><risdate>1993</risdate><volume>122</volume><issue>2</issue><spage>337</spage><epage>347</epage><pages>337-347</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><coden>JCLBA3</coden><abstract>We have mapped the vinculin-binding sites in the cytoskeletal protein talin as well as those sequences which target the talin molecule to focal contacts. Using a series of overlapping talin-fusion proteins expressed in E. coli and (125)I-vinculin in both gel-overlay and microtitre well binding assays, we present evidence for three separable binding sites for vinculin. All three are in the tail segment of talin (residues 434-2541) and are recognized by the same fragment of vinculin (residues 1-258). Two sites are adjacent to each other and span residues 498-950, and the third site is more than 700 residues distant in the primary sequence. Scatchard analysis of (125)I-vinculin binding to talin also indicates three sites, each with a similar affinity (Kd = 2-6 x 10-(7) M). We also detect a substoichiometric interaction of higher affinity (Kd = 3 x 10(-8) M) which remains unexplained. By expressing regions of the chicken talin molecule in heterologous cells, we have shown that the sequences required to target talin to focal contacts overlap those which bind vinculin</abstract><cop>New York, NY</cop><pub>Rockefeller University Press</pub><pmid>8320257</pmid><doi>10.1083/jcb.122.2.337</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0021-9525 |
| ispartof | The Journal of cell biology, 1993-07, Vol.122 (2), p.337-347 |
| issn | 0021-9525 1540-8140 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2119638 |
| source | JSTOR Archival Journals and Primary Sources Collection |
| subjects | 3T3 Cells Animals Binding Sites Biological and medical sciences Cell Line Cell lines Cell structures and functions Cells Cellular biology Chickens Complementary DNA COS cells Cytoskeleton, cytoplasm. Intracellular movements ESTRUCTURA CELULAR Focal adhesions Fundamental and applied biological sciences. Psychology Integrins Mice Molecular and cellular biology Molecules NIH 3T3 cells POLLO POULET Poultry PROTEINAS PROTEINE Proteins Recombinant Fusion Proteins - metabolism STRUCTURE CELLULAIRE Talin - chemistry Talin - genetics Talin - metabolism Transfection Vinculin - metabolism |
| title | The cytoskeletal protein talin contains at least two distinct vinculin binding domains |
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