Skip to main content
Log in

Resonance assignments of the 56 kDa chimeric avidin in the biotin-bound and free forms

  • Article
  • Published:
Biomolecular NMR Assignments Aims and scope Submit manuscript

Abstract

Avidin is a homotetrameric ~56 kDa protein found in chicken egg white. Avidin’s ability to bind biotin with a very high affinity has widely been exploited in biotechnological applications. Protein engineering has further diversified avidin’s feasibility. ChiAVD(I117Y) is a product of rational protein engineering. It is a hyperthermostable synthetic hybrid of avidin and avidin-related protein 4 (AVR4). In this chimeric protein a 23-residue segment in avidin has been replaced with the corresponding sequence found in AVR4, and a point mutation at subunit interface 1–3 (and 2–4) has been introduced. Here we report the backbone and sidechain resonance assignments of the biotin-bound form of ChiAVD(I117Y) as well as the backbone resonance assignments of the free form.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Eisenberg-Domovich Y, Hytönen VP, Wilchek M, Bayer EA, Kulomaa MS, Livnah O (2005) High-resolution crystal structure of an avidin-related protein: insight into high-affinity biotin binding and protein stability. Acta Crystallogr D Biol Crystallogr 61:528–538

    Article  Google Scholar 

  • Heikkinen JJ, Riihimäki TA, Määttä JA, Suomela SE, Kantomaa J, Kulomaa MS, Hytönen VP, Hormi OE (2011) Covalent biofunctionalization of cellulose acetate with thermostable chimeric avidin. ACS Appl Mater Interfaces 3:2240–2245

    Article  Google Scholar 

  • Hytönen VP, Nyholm TK, Pentikäinen OT, Vaarno J, Porkka EJ, Nordlund HR, Johnson MS, Slotte JP, Laitinen OH, Kulomaa MS (2004) Chicken avidin-related protein 4/5 shows superior thermal stability when compared with avidin while retaining high affinity to biotin. J Biol Chem 279:9337–9343

    Article  Google Scholar 

  • Hytönen VP, Määttä JA, Nyholm TK, Livnah O, Eisenberg-Domovich Y, Hyre D, Nordlund HR, Hörhä J, Niskanen EA, Paldanius T, Kulomaa T, Porkka EJ, Stayton PS, Laitinen OH, Kulomaa MS (2005) Design and construction of highly stable, protease resistant chimeric avidins. J Biol Chem 280:10228–10233

    Article  Google Scholar 

  • Laitinen OH, Hytönen VP, Ahlroth MK, Pentikäinen OT, Gallagher C, Nordlund HR, Ovod V, Marttila AT, Porkka E, Heino S, Johnson MS, Airenne KJ, Kulomaa MS (2002) Chicken avidin-related proteins show altered biotin-binding and physico-chemical properties as compared with avidin. Biochem J 363:609–617

    Article  Google Scholar 

  • Laitinen OH, Nordlund HR, Hytönen VP, Kulomaa MS (2007) Brave new (strept) avidins in biotechnology. Trends Biotechnol 25:269–277

    Article  Google Scholar 

  • Livnah O, Bayer EA, Wilchek M, Sussman JL (1993) Three-dimensional structures of avidin and the avidin-biotin complex. Proc Natl Acad Sci USA 90:5076–5080

    Article  ADS  Google Scholar 

  • Määttä JA, Eisenberg-Domovich Y, Nordlund HR, Hayouka R, Kulomaa MS, Livnah O, Hytönen VP (2011) Chimeric avidin shows stability against harsh conditions–biochemical analysis and 3D structure. Biotechnol Bioeng 108:481–490

    Article  Google Scholar 

  • Schwarzinger S, Kroon GJ, Foss TR, Chung J, Wright PE, Dyson HJ (2001) Sequence-dependent correction of random coil NMR chemical shifts. J Am Chem Soc 123:2970–2978

    Article  Google Scholar 

  • Zhang H, Neal S, Wishart DS (2003) RefDB: a database of uniformly referenced protein chemical shifts. J Biomol NMR 25:173–195

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by the Academy of Finland grants 122170 and 131144 to PP.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Perttu Permi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tossavainen, H., Helppolainen, S.H., Määttä, J.A.E. et al. Resonance assignments of the 56 kDa chimeric avidin in the biotin-bound and free forms. Biomol NMR Assign 7, 35–38 (2013). https://doi.org/10.1007/s12104-012-9371-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12104-012-9371-4

Keywords

Navigation