pH-Dependent Deformations of the Energy Landscape of Avidin-like Proteins Investigated by Single Molecule Force Spectroscopy

Melanie Köhler, Andreas Karner, Michael Leitner, Vesa P. Hytönen, Markku Kulomaa, Peter Hinterdorfer, Andreas Ebner

Research output: Contribution to journalReview articlepeer-review

11 Citations (Scopus)


Avidin and avidin-like proteins are widely used in numerous techniques since the avidin-biotin interaction is known to be very robust and reliable. Within this study, we investigated this bond at the molecular level under harsh conditions ranging from very low to very high pH values. We compared avidin with streptavidin and a recently developed avidin-based mutant, chimeric avidin. To gain insights of the energy landscape of these interactions we used a single molecule approach and performed the Single Molecule Force Spectroscopy atomic force microscopy technique. There, the ligand (biotin) is covalently coupled to a sharp AFM tip via a distensible hetero-bi-functional crosslinker, whereas the receptor of interest is immobilized on the probe surface. Receptor-ligand complexes are formed and ruptured by repeatedly approaching and withdrawing the tip from the surface. Varying both pulling velocity and pH value, we could determine changes of the energy landscape of the complexes. Our results clearly demonstrate that avidin, streptavidin and chimeric avidin are stable over a wide pH range although we could identify differences at the outer pH range. Taking this into account, they can be used in a broad range of applications, like surface sensors at extreme pH values.
Original languageEnglish
Pages (from-to)12531-12546
Number of pages16
Issue number8
Publication statusPublished - Aug 2014


  • Avidin mutant
  • Avidin-biotin
  • Biophysics
  • Force spectroscopy
  • Molecular recognition
  • PH dependence
  • Single molecules
  • Immobilized Proteins/chemistry
  • Biotin/chemistry
  • Thermodynamics
  • Streptavidin/chemistry
  • Microscopy, Atomic Force
  • Protein Binding
  • Kinetics
  • Avidin/chemistry
  • Hydrogen-Ion Concentration

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