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Talin and signaling through integrins

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Bouaouina, Mohamed, Harburger, David S., Calderwood, David A.·2011·Methods in Molecular Biology
DOI 10.1007/978-1-61779-166-6_20PMC PMC5642996
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Abstract

Integrin adhesion receptors are essential for the development and functioning of multicellular animals. Integrins mediate cell adhesion to the extracellular matrix and to counter-receptors on adjacent cells, and the ability of integrins to bind extracellular ligands is regulated in response to intracellular signals that act on the short cytoplasmic tails of integrin subunits. Integrin activation, the rapid conversion of integrin receptors from low to high affinity, requires binding of talin to integrin β tails and, once bound, talin provides a connection from activated integrins to the actin cytoskeleton. A wide range of experimental approaches have contributed to the current understanding of the importance of talin in integrin signaling. Here, we describe two methods that have been central to our investigations of talin; a biochemical assay that has allowed characterization of interactions between integrin cytoplasmic tails and talin, and a fluorescent-activated cell-sorting procedure to assess integrin activation in cultured cells expressing talin domains, mutants, dominant negative constructs, or shRNA.

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  • The assays used for integrin activity, LUBAC activity, and ARP2/3 interaction were previously established.

    “As functional read-outs, we used previously established assays for three SHARPIN-regulated functions: integrin activity, LUBAC activity, and ARP2/3 interaction (Bouaouina, Harburger et al., 2011, Harburger, Bouaouina et al., 2009, Khan et al., 2017).”

    From SHARPIN S146 phosphorylation mediates ARP2/3 interaction, cancer cell invasion and metastasis (2022) · cites this paper as Bouaouina, Harburger et al., 2011

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Authors
Bouaouina, M., Harburger, D. S., & Calderwood, D. A.
Affiliations
Yale University (US)
Corresponding author
Calderwood, David A.
ORCID
Calderwood, David A.
Venue
Methods in Molecular Biology
Publisher
Humana Press
Published
2011
Volume
757
Pages
325-347
ISSN
  • 1064-3745 (ISSN-L)
  • 1940-6029
DOI
10.1007/978-1-61779-166-6_20
PMC
PMC5642996
OpenAlex
W204275182
Cited by
42
References
2
Citations by year
2025: 3 · 2023: 1 · 2022: 1 · 2021: 4 · 2020: 4 · 2018: 3 · 2017: 4 · 2016: 4 · 2015: 6 · 2014: 5 · 2013: 3 · 2012: 4
FWCI
6.22
Type
book chapter
Language
en
Open access
Locations
  • Methods in molecular biology (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
National Institute of General Medical Sciences (R01GM088240)
Topics
  • Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
Keywords
talin, integrins, integrin activation, integrin cytoplasmic tail, integrin signaling, cell adhesion, extracellular matrix, actin cytoskeleton, dominant-negative constructs, shRNA knockdown, fluorescence-activated cell sorting, biochemical tests
MeSH terms
Animals, Biological Assay, Flow Cytometry, Cricetinae, Protein Binding, Recombinant Proteins, Signal Transduction, Integrins, CHO Cells, Talin

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