Dimerization of a ubiquitin variant leads to high affinity interactions with a ubiquitin interacting motif

Protein Science : a Publication of the Protein Society
Noah ManczykFrank Sicheri

Abstract

We previously described structural and functional characterization of the first ubiquitin variant (UbV), UbV.v27.1, engineered by phage display to bind with high affinity to a specific ubiquitin interacting motif (UIM). We identified two substitutions relative to ubiquitin (Gly10Val/His68Tyr) that were critical for enhancing binding affinity but could only rationalize the mechanism of action of the Tyr68 substitution. Here, we extend our characterization and uncover the mechanism by which the Val10 substitution enhances binding affinity. We show that Val10 in UbV.v27.1 drives UbV dimerization through an intermolecular β-strand exchange. Dimerization serves to increase the contact surface between the UIM and UbV and also affords direct contacts between two UIMs through an overall 2:2 binding stoichiometry. Our identification of the role of Val10 in UbV dimerization suggests a general means for the development of dimeric UbVs with improved affinity and specificity relative to their monomeric UbV counterparts. Statement: Previously, we used phage display to engineer a UbV that bound tightly and specifically to a UIM. Here, we discovered that tight binding is partly due to the dimerization of the UbV, which increases the contact su...Continue Reading

References

Nov 12, 1991·Biochemistry·H B LowmanJ A Wells
Apr 20, 2002·Science·Titus J BoggonLawrence Shapiro
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Mar 23, 2006·Acta Crystallographica. Section D, Biological Crystallography·Jay Painter, Ethan A Merritt
Aug 2, 2006·Molecular BioSystems·A M Levin, G A Weiss
Aug 8, 2007·Journal of Molecular Biology·Evgeny Krissinel, Kim Henrick
Feb 4, 2010·Acta Crystallographica. Section D, Biological Crystallography·Paul D AdamsPeter H Zwart
Jan 5, 2013·Science·Andreas ErnstSachdev S Sidhu

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Citations

Oct 22, 2019·Journal of Molecular Biology·Pankaj GargSachdev S Sidhu
Jun 13, 2021·Protein Engineering, Design & Selection : PEDS·Chen T LiangWei Zhang

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