SpringerLink
Log in

Studying Protein-Protein Interactions Using Surface Plasmon Resonance

  • Protocol
Protein-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1278))

Abstract

Protein-protein interactions regulate many important cellular processes, including carbohydrate and lipid metabolism, cell cycle and cell death regulation, protein and nucleic acid metabolism, signal transduction, and cellular architecture. A complete understanding of cellular function depends on full characterization of the complex network of cellular protein-protein interactions, including measurements of their kinetic and binding properties. Surface plasmon resonance (SPR) is one of the commonly used technologies for detailed and quantitative studies of protein-protein interactions and determination of their equilibrium and kinetic parameters. SPR provides excellent instrumentation for a label-free, real-time investigation of protein-protein interactions. This chapter details the experimental design and proper use of the instrumentation for a kinetic experiment. It will provide readers with basic theory, assay setup, and the proper way of reporting this type of results with practical tips useful for SPR-based studies. A generic protocol for immobilizing ligands using amino coupling chemistry, also useful if an antibody affinity capture approach is used, performing kinetic studies, and collecting and analyzing data is described.

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

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 179.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Cusick ME, Klitgord N, Vidal M et al (2005) Interactome: gateway into systems biology. Hum Mol Genet 14(Spec No. 2):R171–R181

    Article  CAS  PubMed  Google Scholar 

  2. Ge H, Walhout AJ, Vidal M (2003) Integrating ‘omic’ information: a bridge between genomics and systems biology. Trends Genet 19:551–560

    Article  CAS  PubMed  Google Scholar 

  3. Wells JA, McClendon CL (2007) Reaching for high-hanging fruit in drug discovery at protein-protein interfaces. Nature 450:1001–1009

    Article  CAS  PubMed  Google Scholar 

  4. Blazer LL, Neubig RR (2009) Small molecule protein-protein interaction inhibitors as CNS therapeutic agents: current progress and future hurdles. Neuropsychopharmacology 34:126–141

    Article  CAS  PubMed  Google Scholar 

  5. Ryan DP, Matthews JM (2005) Protein-protein interactions in human disease. Curr Opin Struct Biol 15:441–446

    Article  CAS  PubMed  Google Scholar 

  6. Gerrard JA, Hutton CA, Perugini MA (2007) Inhibiting protein-protein interactions as an emerging paradigm for drug discovery. Mini Rev Med Chem 7:151–157

    Article  CAS  PubMed  Google Scholar 

  7. Ivanov AA, Khuri FR, Fu H (2013) Targeting protein-protein interactions as an anticancer strategy. Trends Pharmacol Sci 34:393–400

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Fagerstam LG, Frostell-Karlsson A, Karlsson R et al (1992) Biospecific interaction analysis using surface plasmon resonance detection applied to kinetic, binding site and concentration analysis. J Chromatogr 597:397–410

    Article  CAS  PubMed  Google Scholar 

  9. Myszka DG (2000) Kinetic, equilibrium, and thermodynamic analysis of macromolecular interactions with BIACORE. Methods Enzymol 323:325–340

    Article  CAS  PubMed  Google Scholar 

  10. Rich RL, Myszka DG (2008) Survey of the year 2007 commercial optical biosensor literature. J Mol Recognit 21:355–400

    Article  CAS  PubMed  Google Scholar 

  11. Hunter MC, O’Hagan KL, Kenyon A et al (2014) Hsp90 binds directly to fibronectin (FN) and inhibition reduces the extracellular fibronectin matrix in breast cancer cells. PLoS One 9:e86842

    Article  PubMed Central  PubMed  Google Scholar 

  12. Chow CR, Suzuki N, Kawamura T et al (2013) Modification of p115RhoGEF Ser(330) regulates its RhoGEF activity. Cell Signal 25:2085–2092

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Pal A, Huang W, Li X et al (2012) CCN6 modulates BMP signaling via the Smad-independent TAK1/p38 pathway, acting to suppress metastasis of breast cancer. Cancer Res 72:4818–4828

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Day YS, Baird CL, Rich RL et al (2002) Direct comparison of binding equilibrium, thermodynamic, and rate constants determined by surface- and solution-based biophysical methods. Protein Sci 11:1017–1025

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Mehand MS, Srinivasan B, De Crescenzo G (2011) Estimation of analyte concentration by surface plasmon resonance-based biosensing using parameter identification techniques. Anal Biochem 419:140–144

    Article  CAS  PubMed  Google Scholar 

  16. Towne V, Zhao Q, Brown M et al (2013) Pairwise antibody footprinting using surface plasmon resonance technology to characterize human papillomavirus type 16 virus-like particles with direct anti-HPV antibody immobilization. J Immunol Methods 388:1–7

    Article  CAS  PubMed  Google Scholar 

  17. Nedelkov D, Nelson RW (2003) Surface plasmon resonance mass spectrometry: recent progress and outlooks. Trends Biotechnol 21:301–305

    Article  CAS  PubMed  Google Scholar 

  18. Williams C, Addona TA (2000) The integration of SPR biosensors with mass spectrometry: possible applications for proteome analysis. Trends Biotechnol 18:45–48

    Article  CAS  PubMed  Google Scholar 

  19. Davis TM, Wilson WD (2000) Determination of the refractive index increments of small molecules for correction of surface plasmon resonance data. Anal Biochem 284:348–353

    Article  CAS  PubMed  Google Scholar 

  20. Shen C, Jo SY, Liao C et al (2013) Targeting recruitment of disruptor of telomeric silencing 1-like (DOT1L): characterizing the interactions between DOT1L and mixed lineage leukemia (MLL) fusion proteins. J Biol Chem 288:30585–30596

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Drake AW, Tang ML, Papalia GA et al (2012) Biacore surface matrix effects on the binding kinetics and affinity of an antigen/antibody complex. Anal Biochem 429:58–69

    Article  CAS  PubMed  Google Scholar 

  22. Pace CN, Vajdos F, Fee L et al (1995) How to measure and predict the molar absorption coefficient of a protein. Protein Sci 4:2411–2423

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Andersson K, Areskoug D, Hardenborg E (1999) Exploring buffer space for molecular interactions. J Mol Recognit 12:310–315

    Article  CAS  PubMed  Google Scholar 

  24. Andersson K, Hamalainen M, Malmqvist M (1999) Identification and optimization of regeneration conditions for affinity-based biosensor assays. A multivariate cocktail approach. Anal Chem 71:2475–2481

    Article  CAS  PubMed  Google Scholar 

  25. Drake AW, Klakamp SL (2011) A strategic and systematic approach for the determination of biosensor regeneration conditions. J Immunol Methods 371:165–169

    Article  CAS  PubMed  Google Scholar 

  26. Karlsson R, Larsson A (2004) Affinity measurement using surface plasmon resonance. Methods Mol Biol 248:389–415

    CAS  PubMed  Google Scholar 

  27. Karlsson R, Katsamba PS, Nordin H et al (2006) Analyzing a kinetic titration series using affinity biosensors. Anal Biochem 349:136–147

    Article  CAS  PubMed  Google Scholar 

  28. Rich RL, Papalia GA, Flynn PJ et al (2009) A global benchmark study using affinity-based biosensors. Anal Biochem 386:194–216

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Cornish-Bowden A (2001) Detection of errors of interpretation in experiments in enzyme kinetics. Methods 24:181–190

    Article  CAS  PubMed  Google Scholar 

  30. Cannon MJ, Papalia GA, Navratilova I et al (2004) Comparative analyses of a small molecule/enzyme interaction by multiple users of Biacore technology. Anal Biochem 330:98–113

    Article  CAS  PubMed  Google Scholar 

  31. Rich RL, Myszka DG (2000) Survey of the 1999 surface plasmon resonance biosensor literature. J Mol Recognit 13:388–407

    Article  CAS  PubMed  Google Scholar 

  32. Rich RL, Myszka DG (2001) Survey of the year 2000 commercial optical biosensor literature. J Mol Recognit 14:273–294

    Article  CAS  PubMed  Google Scholar 

  33. Rich RL, Myszka DG (2002) Survey of the year 2001 commercial optical biosensor literature. J Mol Recognit 15:352–376

    Article  CAS  PubMed  Google Scholar 

  34. Rich RL, Myszka DG (2003) A survey of the year 2002 commercial optical biosensor literature. J Mol Recognit 16:351–382

    Article  CAS  PubMed  Google Scholar 

  35. Rich RL, Myszka DG (2005) Survey of the year 2004 commercial optical biosensor literature. J Mol Recognit 18:431–478

    Article  CAS  PubMed  Google Scholar 

  36. Rich RL, Myszka DG (2005) Survey of the year 2003 commercial optical biosensor literature. J Mol Recognit 18:1–39

    Article  CAS  PubMed  Google Scholar 

  37. Rich RL, Myszka DG (2006) Survey of the year 2005 commercial optical biosensor literature. J Mol Recognit 19:478–534

    Article  CAS  PubMed  Google Scholar 

  38. Rich RL, Myszka DG (2007) Survey of the year 2006 commercial optical biosensor literature. J Mol Recognit 20:300–366

    Article  CAS  PubMed  Google Scholar 

  39. Rich RL, Myszka DG (2010) Grading the commercial optical biosensor literature-Class of 2008: ‘The Mighty Binders’. J Mol Recognit 23:1–64

    Article  CAS  PubMed  Google Scholar 

  40. Katsamba PS, Navratilova I, Calderon-Cacia M et al (2006) Kinetic analysis of a high-affinity antibody/antigen interaction performed by multiple Biacore users. Anal Biochem 352:208–221

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, University of Michigan Medical School, 4510E MSRB I, 1150 West Medical Center Drive, Ann Arbor, MI, 48109, USA

    Zaneta Nikolovska-Coleska

Authors
  1. Zaneta Nikolovska-Coleska
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zaneta Nikolovska-Coleska .

Editor information

Editors and Affiliations

  1. Department of Pharmacology and Emory Chemical Biology Discovery Center, Emory University School of Medicine, Atlanta, Georgia, USA

    Cheryl L. Meyerkord

  2. Department of Pharmacology, Department of Hematology & Medical Oncology, and Emory Chemical Biology Discovery Center, Emory University School of Medicine, Atlanta, USA

    Haian Fu

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Nikolovska-Coleska, Z. (2015). Studying Protein-Protein Interactions Using Surface Plasmon Resonance. In: Meyerkord, C., Fu, H. (eds) Protein-Protein Interactions. Methods in Molecular Biology, vol 1278. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2425-7_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-2425-7_7

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2424-0

  • Online ISBN: 978-1-4939-2425-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation