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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 125I-vinculin in both gel-overlay and microtitre well binding assays,...
Formato: | Texto |
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Lenguaje: | English |
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The Rockefeller University Press
1993
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2119638/ https://www.ncbi.nlm.nih.gov/pubmed/8320257 |
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collection | PubMed |
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 125I-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 125I-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. |
format | Text |
id | pubmed-2119638 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1993 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21196382008-05-01 The cytoskeletal protein talin contains at least two distinct vinculin binding domains J Cell Biol Articles 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 125I-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 125I-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. The Rockefeller University Press 1993-07-02 /pmc/articles/PMC2119638/ /pubmed/8320257 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Articles The cytoskeletal protein talin contains at least two distinct vinculin binding domains |
title | The cytoskeletal protein talin contains at least two distinct vinculin binding domains |
title_full | The cytoskeletal protein talin contains at least two distinct vinculin binding domains |
title_fullStr | The cytoskeletal protein talin contains at least two distinct vinculin binding domains |
title_full_unstemmed | The cytoskeletal protein talin contains at least two distinct vinculin binding domains |
title_short | The cytoskeletal protein talin contains at least two distinct vinculin binding domains |
title_sort | cytoskeletal protein talin contains at least two distinct vinculin binding domains |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2119638/ https://www.ncbi.nlm.nih.gov/pubmed/8320257 |