Skip to main content

Measuring Membrane Protein Interactions Using Optical Biosensors

  • Protocol
  • First Online:
Analgesia

Part of the book series: Methods in Molecular Biology ((MIMB,volume 617))

Abstract

Membrane proteins, such as G protein-coupled receptors (GPCRs) and ion channels, represent important but technically challenging targets for the management of pain and other diseases. Studying their interactions has enabled the development of new therapeutics, diagnostics, and research reagents, but biophysical manipulation of membrane proteins is often difficult because of the requirement of most membrane proteins for an intact lipid bilayer. Here, we describe the use of virus-like particles as presentation vehicles for cellular membrane proteins (“Lipoparticles”). The methods for using Lipoparticles on optical biosensors, such as the BioRad ProteOn XPR36, are discussed as a means to characterize the kinetics, affinity, and specificity of antibody interactions using surface plasmon resonance detection.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Woolf CJ, Ma Q (2007) Nociceptors-noxious stimulus detectors. Neuron 55(3):353–364

    Article  PubMed  CAS  Google Scholar 

  2. Navratilova I, Dioszegi M, Myszka DG (2006) Analyzing ligand and small molecule binding activity of solubilized GPCRs using biosensor technology. Anal Biochem 355(1):132–139

    Article  PubMed  CAS  Google Scholar 

  3. Navratilova I, Sodroski J, Myszka DG (2005) Solubilization, stabilization, and purification of chemokine receptors using biosensor technology. Anal Biochem 339(2):271–281

    Article  PubMed  CAS  Google Scholar 

  4. Rice PJ et al (2002) Human monocyte scavenger receptors are pattern recognition receptors for (1->3)-beta-D-glucans. J Leukoc Biol 72(1):140–146

    PubMed  CAS  Google Scholar 

  5. Stenlund P, Babcock GJ, Sodroski J, Myszka DG (2003) Capture and reconstitution of G protein-coupled receptors on a biosensor surface. Anal Biochem 316(2):243–250

    Article  PubMed  CAS  Google Scholar 

  6. Mobini R et al (2000) A monoclonal antibody directed against an autoimmune epitope on the human beta1-adrenergic receptor recognized in idiopathic dilated cardiomyopathy. . Hybridoma 19(2):135-142 (in eng)

    Article  PubMed  CAS  Google Scholar 

  7. Peter JC, Eftekhari P, Billiald P, Wallukat G, Hoebeke J (2003) scFv single chain antibody variable fragment as inverse agonist of the beta2-adrenergic receptor. (Translated from eng). J Biol Chem 278(38):36740-36747 (in eng)

    Article  PubMed  CAS  Google Scholar 

  8. Day PW et al (2007) A monoclonal antibody for G protein-coupled receptor crystallography. Nat Methods 4(11):927–929

    Article  PubMed  CAS  Google Scholar 

  9. Tummino PJ, Copeland RA (2008) Residence time of receptor-ligand complexes and its effect on biological function. (Translated from eng). Biochemistry 47(20):5481-5492 (in eng)

    Article  PubMed  CAS  Google Scholar 

  10. Canziani G et al (1999) Exploring biomolecular recognition using optical biosensors. (Translated from eng). Methods 19(2):253-269 (in eng)

    Article  PubMed  CAS  Google Scholar 

  11. Willis S et al (2008) Virus-like particles as quantitative probes of membrane protein interactions. Biochemistry 47(27):6988–6990

    Article  PubMed  CAS  Google Scholar 

  12. Bravman T et al (2006) Exploring “one-shot” kinetics and small molecule analysis using the ProteOn XPR36 array biosensor. (Translated from eng). Anal Biochem 358(2):281-288 (in eng)

    Article  PubMed  CAS  Google Scholar 

  13. Hoffman TL, Canziani G, Jia L, Rucker J, Doms RW (2000) A biosensor assay for studying ligand-membrane receptor interactions: binding of antibodies and HIV-1 Env to chemokine receptors. (Translated from eng). Proc Natl Acad Sci USA 97(21):11215-11220 (in eng)

    Article  PubMed  CAS  Google Scholar 

  14. Rucker J (2003) Optical biosensor assay using retroviral receptor pseudotypes. Methods Mol Biol 228:317–328

    PubMed  CAS  Google Scholar 

  15. Karlsson R, Katsamba PS, Nordin H, Pol E, Myszka DG (2006) Analyzing a kinetic titration series using affinity biosensors. (Translated from eng). Anal Biochem 349(1):136-147 (in eng)

    Article  PubMed  CAS  Google Scholar 

  16. Green RJ, Davies MC, Roberts CJ, Tendler SJ (1998) A surface plasmon resonance study of albumin adsorption to PEO-PPO-PEO triblock copolymers. (Translated from eng). J Biomed Mater Res 42(2):165-171 (in eng)

    Article  PubMed  CAS  Google Scholar 

  17. Segura MM et al (2008) Identification of host proteins associated with retroviral vector particles by proteomic analysis of highly purified vector preparations. J Virol 82(3):1107–1117

    Article  PubMed  CAS  Google Scholar 

  18. Myszka DG, He X, Dembo M, Morton TA, Goldstein B (1998) Extending the range of rate constants available from BIACORE: interpreting mass transport-influenced binding data. (Translated from eng). Biophys J 75(2):583-594 (in eng)

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Sharon Willis for her help with Lipoparticle production and biosensor optimization and Laura Moriarty (BioRad) and Mohammed Yousef (BioRad) for their helpful discussions.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Joseph Rucker .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Rucker, J., Davidoff, C., Doranz, B.J. (2010). Measuring Membrane Protein Interactions Using Optical Biosensors. In: Szallasi, A. (eds) Analgesia. Methods in Molecular Biology, vol 617. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-323-7_32

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-323-7_32

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-322-0

  • Online ISBN: 978-1-60327-323-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics