An Oncometabolite Isomer Rapidly Induces a Pathophysiological Protein Modification.

Area of Science:

• Biochemistry and Molecular Biology
• Chemical Biology
• Cellular Signaling

Background:

• Metabolites modulate protein activity through covalent and noncovalent interactions.
• Controlling metabolite-protein interactions within living cells presents significant challenges.

Purpose of the Study:

• To develop a chemical strategy for inducing cysteine S-succination, a nonenzymatic post-translational modification.
• To evaluate novel S-succination inducers, maleate and 2-bromosuccinate, for their reactivity and cellular effects.
• To investigate the potential of induced S-succination in modulating cellular signaling pathways, specifically KEAP1-NRF2.

Main Methods:

• Utilized antibody-based detection for monitoring S-succination.
• Employed kinetic assays to assess the reactivity of S-succination inducers.
• Conducted cellular assays to evaluate the efficacy and toxicity of maleate and 2-bromosuccinate in living cells.

Main Results:

• Identified maleate and 2-bromosuccinate as novel chemical inducers of cysteine S-succination.
• Demonstrated that maleate is a more potent and less toxic inducer of S-succination in cellular contexts.
• Showed that maleate-induced S-succination activates the KEAP1-NRF2 signaling pathway in living cells.

Conclusions:

• Established a chemical method to induce cysteine S-succination, mimicking an oncometabolite-protein interaction.
• Provided a robust platform for studying metabolite-protein interactions with high physiochemical accuracy and low toxicity.
• This approach offers a methodological foundation for understanding the role of metabolites in disease signaling.