High-affinity binders selected from designed ankyrin repeat protein libraries

@article{Binz2004HighaffinityBS,
  title={High-affinity binders selected from designed ankyrin repeat protein libraries},
  author={H. Kaspar Binz and Patrick Amstutz and Andreas Kohl and Michael T. Stumpp and Christophe Briand and Patrik Forrer and Markus G. Grütter and Andreas Pl{\"u}ckthun},
  journal={Nature Biotechnology},
  year={2004},
  volume={22},
  pages={575-582}
}
We report here the evolution of ankyrin repeat (AR) proteins in vitro for specific, high-affinity target binding. Using a consensus design strategy, we generated combinatorial libraries of AR proteins of varying repeat numbers with diversified binding surfaces. Libraries of two and three repeats, flanked by 'capping repeats,' were used in ribosome-display selections against maltose binding protein (MBP) and two eukaryotic kinases. We rapidly enriched target-specific binders with affinities in… 
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TLDR
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Destabilizing an interacting motif strengthens the association of a designed ankyrin repeat protein with tubulin
TLDR
Biochemical and structural characterizations demonstrated that the evolved mutants achieved a gain in affinity through destabilization of the C-cap, which relieves the need of a DARPin conformational change upon tubulin binding and removes unfavorable interactions in the complex.
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