Related Experiment Video

Updated: Oct 23, 2025

Evaluation of Substrate Ubiquitylation by E3 Ubiquitin-ligase in Mammalian Cell Lysates
09:47

Evaluation of Substrate Ubiquitylation by E3 Ubiquitin-ligase in Mammalian Cell Lysates

Published on: May 10, 2022

2.8K

Inducing Ubiquitylation and Protein Degradation as a Drug Development Strategy

Kamaldeep S Dhami1, XiaoDong Huang2

  • 1Oncology Discovery, AbbVie, Sunnyvale, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 25, 2021
PubMed

Insights

New drug discovery methods use the ubiquitin-proteasome system (UPS) to degrade oncoproteins. Protocols are presented to assess protein ubiquitination, degrader-induced degradation, and cancer cell proliferation for evaluating drug targets.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Drug discovery increasingly utilizes protein degradation via the ubiquitin-proteasome system (UPS).
  • Established degrader platforms include IMiDs, PROTACs (proteolysis targeting chimeras), and molecular glues, with several in clinical development.
  • Protein degraders offer advantages over inhibitors, including catalytic mechanisms and sustained efficacy.

Purpose of the Study:

  • To present standardized protocols for evaluating drug targets using degrader platforms.
  • To enable measurement of intrinsic protein ubiquitination.
  • To quantify degrader-induced target protein degradation and assess cancer cell proliferation.

Main Methods:

  • Measurement of intrinsic protein ubiquitination levels.
  • Quantification of target protein degradation induced by small molecule degraders.
  • Assessment of cancer cell proliferation in response to degrader treatment.

Main Results:

  • Established protocols for evaluating protein ubiquitination and degradation.
  • Demonstrated methods for assessing the impact of degraders on cancer cell proliferation.
  • Provided a framework for assessing the potential of drug targets with degrader platforms.

Conclusions:

  • The described protocols facilitate the evaluation of drug targets for degrader-based therapies.
  • These methods are crucial for advancing the development of novel oncoprotein degradation strategies.
  • Standardized assessment of protein degradation and cellular effects is key for drug discovery.
Keywords:
DegraderInhibitorPROTACProteasomeUbiquitin

More Related Videos

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases06:30

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases

Published on: August 27, 2021

3.3K
High-Throughput Cellular Profiling of Targeted Protein Degradation Compounds Using HiBiT CRISPR Cell Lines05:33

High-Throughput Cellular Profiling of Targeted Protein Degradation Compounds Using HiBiT CRISPR Cell Lines

Published on: November 9, 2020

10.3K

Related Concept Videos

Regulated Protein Degradation02:58

Regulated Protein Degradation

It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
7.9K
The Proteasome01:13

The Proteasome

Eukaryotic cells can degrade proteins through several pathways. One of the most important among these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
1.2K
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
8.0K
Receptor Downregulation in MVBs01:15

Receptor Downregulation in MVBs

Multivesicular bodies (MVBs) are mature endosomes that sort ubiquitinated proteins and then fuse with lysosomes to degrade the sorted proteins. Epidermal growth factor (EGF) and its receptor (EGFR) form a complex that can be internalized through endocytosis, sorted into an MVB, and later degraded.
The EGFR can initiate signaling pathways that  lead to cell proliferation, migration, and differentiation. Overexpression of EGFR  stimulates cells to proliferate. Excessive  EGFR...
2.3K
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
1.1K
The Unfolded Protein Response01:37

The Unfolded Protein Response

The ER is the hub of protein synthesis in a cell. It has robust systems to quality control protein folding and also for degradation of terminally misfolded proteins. Under normal conditions, a small proportion of misfolded proteins that cannot be salvaged need to be transported to the cytoplasm by the ER-associated degradation or ERAD pathways. However, if the ERAD cannot handle the misfolded proteins, the cell activates the unfolded protein response or UPR to adjust the protein folding...
5.5K