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Biochemical characterization of the Integrin Interactome.

Khan, Rejina B. and Varela, Lorena and Cowell, Alana Romain and Goult, Benjamin T (2020) Biochemical characterization of the Integrin Interactome. In: The Integrin Interactome - Methods and Protocols. Methods in Molecular Biology . Springer Protocols, pp. 115-147. ISBN 978-1-07-160962-0. (doi:10.1007/978-1-0716-0962-0_9) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:85472)

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More than 250 proteins are associated with the formation of integrin adhesion complexes involving a vast number of complex interactions between them. These interactions enable adhesions to serve as dynamic and diverse mechanosignaling centers. Our laboratory focuses on the biochemical and structural study of these interactions to help unpick this complex network. Here, we describe the general pipeline of biochem- ical assays and methods we use. The chapter is split into two sections: (1) protein production and characterization and (2) biochemical assays for the characterization of binding between full-length proteins and/or specific regions of proteins with other proteins, peptides, and phospholipids. The suite of assays we use routinely includes circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy for sample quality assessment, prior to biochemical analysis using NMR, fluorescence polarization (FP), microscale thermophoresis (MST), size-exclusion chromatography multiangle light scattering (SEC-MALS), and pulldown/cosedimentation-based approaches. The results of our analysis feed into in vivo studies that allow for the elucidation of the biological role of each interaction.

Item Type: Book section
DOI/Identification number: 10.1007/978-1-0716-0962-0_9
Uncontrolled keywords: Integrin, Adhesome, Interaction, Biochemistry, talin
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Ben Goult
Date Deposited: 12 Jan 2021 20:22 UTC
Last Modified: 14 Nov 2022 23:11 UTC
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