3FWV

Crystal Structure of a Redesigned TPR Protein, T-MOD(VMY), in Complex with MEEVF Peptide

Summary for 3FWV

• Entry DOI: 10.2210/pdb3fwv/pdb
• Descriptor: Hsc70/Hsp90-organizing protein, Heat shock protein HSP 90-beta, NICKEL (II) ION, ... (4 entities in total)
• Functional Keywords: tetratricopeptide repeat protein (tpr), protein-peptide complex, designed protein, tpr, nucleus, phosphoprotein, tpr repeat, chaperone
• Biological source: Homo sapiens (human); More
• Cellular location: Cytoplasm (By similarity): P31948; Cytoplasm: P08238
• Total number of polymer chains: 4
• Total formula weight: 32002.96

Authors

Jackrel, M.E.,Valverde, R.,Regan, L. (deposition date: 2009-01-19, release date: 2009-04-21, Last modification date: 2021-10-20)

Primary citation

Jackrel, M.E.,Valverde, R.,Regan, L.
Redesign of a protein-peptide interaction: characterization and applications
Protein Sci., 18:762-774, 2009

PubMed Abstract: 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.

PubMed: 19309728
DOI: 10.1002/pro.75

Experimental method

X-RAY DIFFRACTION (2.2 Å)