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PMID: 1694125 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Antibody mapping of functional domains in vinculin.

The EMBO journal ·Vol. 9 ·No. 7 ·1990-07-00 ·Pages 2071-8

Westmeyer A, Ruhnau K, Wegner A, Jockusch BM

Abstract

We have analyzed the functional domain structure of vinculin, a protein involved in linking microfilaments to the cytoplasmic face of cell membranes in animal cells. For this purpose, we used several monoclonal antibodies raised against chicken gizzard vinculin whose epitopes could be assigned to discrete regions in the vinculin sequence by immunoblotting of proteolytic fragments combined with N-terminal amino acid sequencing. Two of these antibodies induced the disruption of stress fibers and changed the number of morphology of focal contacts after microinjection in chicken embryo fibroblasts. Based on the location of its epitope in comparison with vinculin domains previously identified by other groups, we propose that one of these antibodies (15B7) interferes with the binding of vinculin to talin, the most peripheral of the microfilament proteins. The second antibody (14C10) binds within a region comprising three internal repeats and might therefore distort the inner architecture of vinculin. A third antibody (As3) inhibited the binding of F-actin to vinculin in an in vitro assay but had no effect on the microfilament system in cells. These data emphasize the role of vinculin as a key protein in microfilament-membrane linkage and support previous work on a direct interaction between vinculin and actin.

MeSH Terms
Actins/metabolism Animals Antibodies, Monoclonal Antigen-Antibody Complex Binding Sites Chick Embryo Chickens Cytoskeletal Proteins/genetics,immunology,metabolism Enzyme-Linked Immunosorbent Assay Epitopes/analysis Fibroblasts/metabolism Gizzard, Avian/metabolism Immunoblotting Immunoglobulin G Muscle, Smooth/metabolism Peptide Fragments/isolation & purification Restriction Mapping Vinculin
Chemicals
Actins Antibodies, Monoclonal Antigen-Antibody Complex Cytoskeletal Proteins Epitopes Immunoglobulin G Peptide Fragments Vinculin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Westmeyer A
Developmental Biology Unit, University of Bielefeld, FRG.
Ruhnau K
Wegner A
Jockusch B M
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54 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-07-00
Pages
2071-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC551924
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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