Design and Functional Characterization of Synthetic E3 Ubiquitin Ligases for Targeted Protein Depletion. Issue 1 (9th April 2018)

• Record Type: Journal Article
• Authors: Baltz, Morgan R.; Stephens, Erin A.; DeLisa, Matthew P.
• Editors: Mahal, Lara; Romesberg, Floyd; Shah, Kavita; Shamu, Caroline; Strano, Michael S.; Thomas, Craig
• Is Part Of: Current protocols in chemical biology. Volume 10:Issue 1(2018)
• Page Start: 72
• Page End: 90
• Publication Date: 2018-04-09
• Subjects: antibody -- non‐antibody protein scaffolds -- protein degradation -- protein silencing -- ubiquibody -- ubiquitination -- ubiquitin proteasome pathway (UPP); Biochemistry -- Laboratory manuals; Chemical Phenomena; Biochemistry; Periodicals; Laboratory manuals; 572.078
• DOI: 10.1002/cpch.37
• Languages: English
• ISSNs: 2160-4762

Abstract

Abstract: A number of techniques now exist for decreasing the expression of cellular proteins without the need for genomic modification. One such technique involves engineered protein chimeras that combine the ubiquitination activity of E3 ubiquitin ligases with the binding affinity and substrate specificity of designer binding proteins (DBPs). These chimeras, called "ubiquibodies, " are capable of selectively and controllably steering virtually any protein to the ubiquitin proteasome pathway (UPP) for degradation, making ubiquibodies a powerful addition to the protein knockout toolbox. A distinguishing feature of ubiquibodies is their modularity—simply swapping DBPs can generate a new ubiquibody with specificity for a different substrate protein. Moreover, by employing DBPs that recognize particular protein states (e.g., active versus inactive conformation, mutant versus wild‐type, post‐translational modification), it becomes possible to deplete certain protein subpopulations while sparing others. This protocol outlines the steps necessary to design and functionally evaluate ubiquibodies for customizable silencing of cellular proteins. © 2018 by John Wiley & Sons, Inc.