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PDBsum entry 3fwv
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References listed in PDB file
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Key reference
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Title
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Redesign of a protein-Peptide interaction: characterization and applications.
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Authors
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M.E.Jackrel,
R.Valverde,
L.Regan.
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Ref.
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Protein Sci, 2009,
18,
762-774.
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PubMed id
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Abstract
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The design of protein-peptide interactions has a wide array of practical
applications and also reveals insight into the basis for molecular recognition.
Here, we present the redesign of a tetratricopeptide repeat (TPR) protein
scaffold, along with its corresponding peptide ligand. We show that the binding
properties of these protein-peptide pairs can be understood, quantitatively,
using straightforward chemical considerations. The recognition pairs we have
developed are also practically useful for the specific identification of tagged
proteins. We demonstrate the facile replacement of these proteins, which we have
termed T-Mods (TPR-based recognition module), for antibodies in both detection
and purification applications. The new protein-peptide pair has a dissociation
constant that is weaker than typical antibody-antigen interactions, yet the
recognition pair is highly specific and we have shown that this affinity is
sufficient for both Western blotting and affinity purification. Moreover, we
demonstrate that this more moderate affinity is actually advantageous for
purification applications, because extremely harsh conditions are not required
to dissociate the T-Mod-peptide interaction. The results we present are
important, not only because they represent a successful application of protein
design but also because they help define the properties that should be sought in
other scaffolds that are being developed as antibody replacements.
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