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Thermal Proteome Profiling Reveals the O-GlcNAc-Dependent Meltome
Dustin T King1,2, Jesús E Serrano-Negrón1, Yanping Zhu1,2
1Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, British Columbia V5A 1S6, Canada.
Journal of the American Chemical Society
|March 1, 2022
Summary
Posttranslational modifications impact protein stability. A new thermal proteomic profiling method reveals O-linked N-acetylglucosamine (O-GlcNAc) often destabilizes proteins, challenging previous assumptions and showing bidirectional regulation.
Area of Science:
- Biochemistry
- Proteomics
- Cellular Biology
Background:
- Posttranslational modifications (PTMs) are crucial for protein function and cellular physiology.
- PTMs can alter protein stability, influencing cellular processes.
- Global methods to assess PTM effects on proteome stability are limited.
Purpose of the Study:
- To develop and apply an unbiased, proteome-wide approach to investigate the impact of PTMs on protein thermodynamic stability.
- To explore the specific effects of O-linked N-acetylglucosamine (O-GlcNAc) on protein stability across the proteome.
Main Methods:
- Developed a thermal proteomic profiling strategy for large-scale assessment of protein stability.
- Applied the method to study O-GlcNAc modification in mammalian systems.
- Validated findings using orthogonal biochemical and proteomic techniques.
Main Results:
- Identified 72 proteins exhibiting O-GlcNAc-dependent changes in thermostability.
- Contrary to expectations, the majority of O-GlcNAc-modified proteins were destabilized.
- Affected proteins were found to be enriched in specific macromolecular complexes.
Conclusions:
- O-linked N-acetylglucosamine (O-GlcNAc) acts as a bidirectional regulator of protein stability.
- The developed thermal proteomic profiling strategy provides a framework for studying PTM impacts on protein stability.
- This approach can be broadly applied to investigate any PTM in any organism.
