The Integrin Interactome, 1st ed. 2021 Methods and Protocols Methods in Molecular Biology Series, Vol. 2217

This volume provides the most cutting edge technologies related to the study of integrin activation and the characterization of their vast interactomes. Chapters detail protocols on experimental approached to quantify focal adhesion parameters, integrin activation, and the lateral interaction of integrins with transmembrane binding partners. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, The Integrin Interactome: Methods and Protocols aims to give the reader a multi-scale journey from single bonds inside protein structures to the function of these crucial adhesion receptors at a whole organism level in physiology and pathology.

1. Measurement of Integrin Activation and Conformational Changes on the Cell Surface by Soluble Ligand and Antibody Binding Assays

Zhengli Wang and Jieqing Zhu

2. Quantification of Integrin Activation and Ligation in Adherent Cells

Zaki Al-Yafeai and A. Wayne Orr

3. Multi-parametric Analysis of Focal Adhesions in Bi-Dimensional Substrates

Vanessa C. Talayero and Miguel Vicente-Manzanares

4. Focal Adhesion Isolation Assay using ECM Coated Magnetic Beads

Wesley Sturgess and Vinay Swaminathan

5. Functional Integrin Regulation through Interactions with Tetraspanin CD9

Álvaro Torres-Gómez, Beatriz Cardeñes, Ester Díez-Sainz, Esther M. Lafuente, and Carlos Cabañas

6. Proximity-dependent Biotinylation (BioID) of Integrin Interaction Partners

Satu-Marja Myllymäki, Xiaonan Liu, Markku Varjosalo, and Aki Manninen

7. Analyzing the Integrin Adhesome by in situ Proximity Ligation Assay

Brian A. Perrino, Yeming Xie, and Cristina Alexandru

8. Single-Protein Tracking to Study Protein Interactions during integrin-based Migration

A.V. Radhakrishnan, Tianchi Chen, Filipe Nunes Vicente, Thomas Orré, Amine Mehidi, Olivier Rossier, and Grégory Giannone

9. Biochemical Characterization of the Integrin Interactome

Rejina B. Khan, Lorena Varela, Alana R. Cowell, and Benjamin T. Goult

10. Network Analysis of Integrin Adhesion Complexes

Frederic Li Mow Chee and Adam Byron

11.       Surface Patterning for the Control of Receptor Clustering and Molecular Forces of Integrin-mediated Adhesions

Federica Pennarola and E. Ada Cavalcanti-Adam

12. Dynamics and Physics of Integrin Activation in Tumour Cells by Nano-Sized Extracellular Ligands and Electromagnetic Fields 

Alkiviadis-Constantinos Cefalas, Vassilios Gavriil, Angelo Ferraro, Zoe Kollia, and Evangelia Sarantopoulou

13. Genetic Instruction of Megakaryocytes and Platelets Derived from Human Induced Pluripotent Stem Cells for Studies of Integrin Regulation

Ana Kasirer-Friede and Sanford J. Shattil

14. Quantitative Analysis of Integrin Trafficking

Enoir Farage and Patrick T. Caswell

15. Methods to Study Integrin Functions on Exosomes

Eiji Kawamoto, Eun Jeong Park, and Motomu Shimaoka

Edward Roy Horton

17. Quantifying Polarized Extracellular Matrix Secretion in Cultured Endothelial Cells

Fabiana Clapero, Dora Tortarolo, Donatella Valdembri, and Guido Serini

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts