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Unraveling Ribonucleic Acid Unfolding: A Quantitative Comparison of Solution and Gas-Phase Unfolding
Anna G Anders1, Jacqueline Anthenien1, Ingrid R Kilde2
1Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States.
Analytical Chemistry
|May 23, 2025
Summary
Collision-induced unfolding (CIU) advancements now quantitatively link gas-phase RNA structures to solution-phase data. This improved method, applied to MELAS-associated mt-tRNA, reveals strong correlations relevant to RNA
Area of Science:
- Structural Biology
- Biophysics
- Molecular Biology
Background:
- Ribonucleic acids (RNAs) present significant challenges for structural biology studies.
- Collision-induced unfolding (CIU) offers a rapid method for assessing noncoding RNA higher-order structure.
- Current correlations between CIU data and solution structures are largely qualitative.
Purpose of the Study:
- To advance RNA collision-induced unfolding (CIU) methods.
- To quantitatively correlate gas-phase CIU data with solution-phase unfolding data.
- To apply enhanced CIU to disease-associated RNA, specifically MELAS-associated mt-tRNA^Leu(UUR).
Main Methods:
- Implemented "supercharging" to enhance RNA CIU.
- Quantitatively evaluated supercharged RNA CIU data.
- Collected and compared solution-phase unfolding data across varying Mg2+ concentrations.
- Applied supercharged CIU to MELAS-associated mt-tRNA^Leu(UUR).
Main Results:
- Supercharging significantly improved information obtained from RNA CIU.
- Demonstrated strong quantitative correlations between gas-phase CIU and solution-phase unfolding data.
- Observed correlations were dependent on Mg2+ concentration and MELAS-associated mutations.
- Established solution-relevant relationships for CIU data in RNAs.
Conclusions:
- Advanced RNA CIU provides quantitative, solution-relevant structural insights.
- CIU is a powerful tool for biophysical characterization of disease-associated RNAs.
- Future work includes CIU annotation, broader biophysical studies, and transcriptomic analysis.
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