Resonance assignments of the 56 kDa chimeric avidin in the biotin-bound and free forms
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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.
KeywordsBiotin Chimeric avidin NMR Thermostability Ligand binding
- 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 279:9337–9343CrossRefGoogle 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 280:10228–10233CrossRefGoogle 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 363:609–617CrossRefGoogle Scholar