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Related Experiment Video

Updated: Jul 6, 2026

NMR 15N Relaxation Experiments for the Investigation of Picosecond to Nanoseconds Structural Dynamics of Proteins

NMR ¹⁵N Relaxation Experiments for the Investigation of Picosecond to Nanoseconds Structural Dynamics of Proteins

Published on: November 1, 2024

Mapping functional dynamics hotspots for protein engineering with NMR peak intensity analysis.

Adam M Damry¹, Serena E Hunt¹, Sandrine Legault¹

¹Department of Chemistry and Biomolecular Sciences, University of Ottawa, 10 Marie-Curie, Ottawa, ON K1N 6N5, Canada.

Protein Engineering, Design & Selection : PEDS

|July 5, 2026

Summary

Nuclear Magnetic Resonance (NMR) peak intensity analysis quickly identifies protein dynamics hotspots. This method guides protein engineering by revealing how structural flexibility impacts protein function, enhancing activity.

Keywords:

NMR X-ray crystallography fluorescent proteins protein dynamics protein engineering

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Last Updated: Jul 6, 2026

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Measuring Interactions of Globular and Filamentous Proteins by Nuclear Magnetic Resonance Spectroscopy (NMR) and Microscale Thermophoresis (MST)

Measuring Interactions of Globular and Filamentous Proteins by Nuclear Magnetic Resonance Spectroscopy (NMR) and Microscale Thermophoresis (MST)

Published on: November 2, 2018

Area of Science:

Biochemistry and Structural Biology
Protein Engineering and Design
Biophysical Chemistry

Background:

Protein structural dynamics are critical for biological function.
Protein engineering aims to enhance protein activity by modifying dynamics.
Identifying dynamic hotspots for mutagenesis is currently challenging and time-consuming.

Purpose of the Study:

To establish Nuclear Magnetic Resonance (NMR) peak intensity analysis as a rapid and precise method for identifying functionally relevant protein dynamics hotspots.
To guide protein engineering strategies by mapping dynamic regions correlated with protein function.

Main Methods:

Utilized NMR peak intensity analysis, a residue-level resolution technique, to assess protein flexibility.
Studied a family of red fluorescent proteins (RFPs) as a model system.
Corroborated findings using B-factor analysis from non-cryogenic X-ray crystal structures.

Main Results:

NMR peak intensity analysis effectively identified functionally relevant dynamic regions in RFPs.
Observed a correlation between structural flexibility and quantum yield in RFPs.
Demonstrated that increased quantum yield leads to rigidity near the chromophore phenolate and flexibility near the acylimine group, with the phenolate face being a known mutational hotspot.

Conclusions:

NMR peak intensity analysis is a rapid, high-precision tool for mapping protein dynamics hotspots.
This method can effectively guide protein engineering efforts aimed at enhancing protein function and activity.
The identified dynamic regions and their correlation with function provide insights into RFP engineering.