Journal
/
Biochemistry
/
Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases

DOI:
10.3791/62950-v

•

06:30 min

•

August 27, 2021

•

,
1Department of Molecular and Cellular Biology, College of Biological Science,University of Guelph

Chapters

  • 00:04Introduction
  • 00:36Round One of Selection
  • 03:23Preparation for the Subsequent Rounds of Selection
  • 04:37Results: Ubiquitin Variant (UbV) Selection Against UBE4B
  • 05:43Conclusion

Summary

Automatic Translation

Automatic Translation

Ubiquitination is a critical protein post-translational modification, dysregulation of which has been implicated in numerous human diseases. This protocol details how phage display can be utilized to isolate novel ubiquitin variants that can bind and modulate the activity of E3 ligases that control the specificity, efficiency, and patterns of ubiquitination.

Tags

Keywords: Phage DisplayUbiquitin VariantsE3 LigasesModulatorsBindersTarget ProteinsAffinity SelectionPanningElution

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/62950?article_type=v

Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
check_url/62950?article_type=v

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro
check_url/62950?article_type=v

Semi-automated Biopanning of Bacterial Display Libraries for Peptide Affinity Reagent Discovery and Analysis of Resulting Isolates
check_url/62950?article_type=v

An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
check_url/62950?article_type=v

Dissipative Microgravimetry to Study the Binding Dynamics of the Phospholipid Binding Protein Annexin A2 to Solid-supported Lipid Bilayers Using a Quartz Resonator
check_url/62950?article_type=v

Using High Content Imaging to Quantify Target Engagement in Adherent Cells
check_url/62950?article_type=v

Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker
check_url/62950?article_type=v

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale
check_url/62950?article_type=v

Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations
check_url/62950?article_type=v

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Read Article