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Surface Plasmon Resonance Biosensing in Studies of the Binding Between β2 Integrin I Domains and Their Ligands

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Integrin and Cell Adhesion Molecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 757))

Abstract

Measurements on the kinetic aspects of binding between macromolecular species such as proteins have been greatly advanced by the application of surface plasmon resonance (SPR) biosensors. In studies of ligand binding by integrin I domains, technologies such as the BIAcore instruments have provided important insights into the role of conformational regulation. This chapter describes a protocol for studying the binding between the I domain from integrin αXβ2 and its ligand iC3b. Also included are topics on the interpretation of data. Integrin I domains appear to support heterogeneous interactions with ligands, which pose significant challenges in deriving valid information on the binding kinetics from the SPR measurements. Fortunately, new algorithms are available that may resolve even complex ligand-binding reactions; with the application to data on the binding between the αX I domain, a more consistent and unambiguous result is obtained compared to those obtained by classical approaches for analyzing SPR biosensor data.

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Acknowledgments

I am indebted to Drs. Timothy A. Springer and Motomu Shimaoka for introducing me to integrin biology and the details of SPR instrumentation. Likewise, I thank Dr. Junichi Takagi for several helpful suggestions and intellectual support on the experimental investigations of integrin–ligand binding. Drs. Peter Schuck and Juraj Svitel kindly provided their MatLab code for SPR data analysis, and helped the initial application of this approach to our data. Dr. Malcolm W. Turner kindly helped the writing of this chapter. This work was supported by grants from the Carlsberg Foundation and LEO Pharma Research Foundation.

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Authors and Affiliations

  1. Biophysical Immunology Laboratory, Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus C, Denmark

    Thomas Vorup-Jensen

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  1. Thomas Vorup-Jensen
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Correspondence to Thomas Vorup-Jensen .

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Editors and Affiliations

  1. Immune Disease Institute, Harvard Medical School, Longwood Avenue 200, Boston, 02115, Massachusetts, USA

    Motomu Shimaoka

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Vorup-Jensen, T. (2011). Surface Plasmon Resonance Biosensing in Studies of the Binding Between β2 Integrin I Domains and Their Ligands. In: Shimaoka, M. (eds) Integrin and Cell Adhesion Molecules. Methods in Molecular Biology, vol 757. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-166-6_5

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  • DOI: https://doi.org/10.1007/978-1-61779-166-6_5

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-165-9

  • Online ISBN: 978-1-61779-166-6

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