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Related Experiment Video

Updated: May 30, 2026

A Semi-Quantitative Drug Affinity Responsive Target Stability (DARTS) assay for studying Rapamycin/mTOR interaction

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Published on: August 27, 2019

Proteomic analysis shows synthetic oleanane triterpenoid binds to mTOR.

Mark M Yore¹, Arminja N Kettenbach, Michael B Sporn

¹Department of Pharmacology, Dartmouth Medical School, Hanover, New Hampshire, United States of America.

|August 6, 2011

Summary

New synthetic oleanane triterpenoids (SO) like CDDO-Imidazolide (CDDO-Im) target multiple cellular networks. This study identifies 577 binding proteins, including mTOR, revealing CDDO-Im

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Published on: October 23, 2018

Area of Science:

Pharmacology
Proteomics
Molecular Biology

Background:

Multifunctional drugs targeting disease networks offer novel therapeutic strategies.
Synthetic oleanane triterpenoids (SO) are developed for inflammation and oxidative stress but their targets are not fully understood.

Purpose of the Study:

To characterize the protein binding partners and mechanisms of action of synthetic oleanane triterpenoid CDDO-Imidazolide (CDDO-Im).

Main Methods:

Affinity purification coupled with mass spectrometry to identify CDDO-Im binding proteins.
Bioinformatic analysis of the resulting protein interactome.
Pull-down assays to validate specific targets.
Biochemical assays to assess mTOR kinase activity.

Main Results:

Identified 577 candidate binding proteins for CDDO-Im, forming interconnected signaling networks.
Bioinformatic analysis revealed targeted pathways including retinoic acid receptor (RAR), estrogen receptor (ER), insulin receptor (IR), JAK/STAT, and PTEN.
Demonstrated that mammalian target of rapamycin (mTOR) is a direct target of CDDO-Im.
Showed CDDO-Im inhibits insulin-induced mTOR activation by binding to and reducing its kinase activity.

Conclusions:

CDDO-Im acts on a complex protein network to exert its pharmacological effects.
mTOR is a direct target of CDDO-Im, and its inhibition contributes to the drug's mechanism of action.
These findings advance the understanding of SO mechanisms and their therapeutic potential.