Related Experiment Video
Updated: Sep 23, 2025
10:28
Measuring Interactions of Globular and Filamentous Proteins by Nuclear Magnetic Resonance Spectroscopy NMR and Microscale Thermophoresis MST
Published on: November 2, 2018
12.3K
19F multiple-quantum coherence NMR spectroscopy for probing protein-ligand interactions
Anna Zawadzka-Kazimierczuk1,2, Mate Somlyay1, Hanspeter Kaehlig3
1Department of Structural and Computational Biology, Max F. Perutz Laboratories, University of Vienna Vienna Biocenter Campus 5 A-1030 Vienna Austria.
RSC Advances
|May 13, 2022
Summary
A novel fluorine-19 NMR technique identifies weak protein binders using unique SF5 compounds and five-quantum coherences (5QC). This method aids early drug discovery by detecting low-affinity interactions with targets like NGAL and beta-catenin.
Area of Science:
- Biophysical Chemistry
- Structural Biology
- Drug Discovery
Background:
- Detecting weak molecular interactions is crucial for early drug discovery.
- Existing methods may lack sensitivity for identifying low-affinity binders.
- Fluorine-19 Nuclear Magnetic Resonance (19F NMR) offers unique advantages for studying biomolecular interactions.
Purpose of the Study:
- To introduce a novel 19F NMR method for detecting weak protein-small molecule binding (up to mM affinity).
- To leverage synthetic SF5-containing compounds and five-quantum coherences (5QC) for enhanced sensitivity.
- To demonstrate the method's utility in early-stage drug discovery programs.
Main Methods:
- Utilizing synthetic compounds containing the pentafluorosulfanyl (SF5) group.
- Generating and analyzing five-quantum coherences (5QC) in 19F NMR.
- Monitoring 5QC relaxation changes indicative of reversible binding events.
Main Results:
- The 5QC relaxation is highly sensitive to binding-induced exchange events.
- Weakly binding fragments can be reliably identified using this approach.
- Successful application demonstrated on neurotoxic NGAL and tumor target β-catenin.
Conclusions:
- This new 19F NMR method provides a sensitive tool for detecting weak protein-ligand interactions.
- It is particularly valuable for fragment-based drug discovery and lead identification.
- The technique expands the capabilities of NMR in characterizing early-stage drug candidates.
Related Concept Videos
01:182D NMR: Overview of Heteronuclear Correlation Techniques
328
Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an X-nucleus, such as carbon-13 or nitrogen-15. This method is commonly used in nuclear magnetic resonance (NMR) spectroscopy to gain insights into complex chemical compounds' structural and compositional aspects. A typical heteronuclear correlation spectrum displays X-nucleus chemical shifts on one axis and a proton spectrum on the other...
328
01:192D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)
948
Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does...
948
01:08NMR Spectroscopy: Spin–Spin Coupling
1.7K
The spin state of an NMR-active nucleus can have a slight effect on its immediate electronic environment. This effect propagates through the intervening bonds and affects the electronic environments of NMR-active nuclei up to three bonds away; occasionally, even farther. This phenomenon is called spin–spin coupling or J-coupling. Coupling interactions are mutual and result in small changes in the absorption frequencies of both nuclei involved. While nuclei of the same element are involved...
1.7K
01:162D NMR: Overview of Homonuclear Correlation Techniques
305
Homonuclear correlation spectroscopy (COSY) is a powerful technique used in Nuclear Magnetic Resonance (NMR) spectroscopy to study the correlations between nuclei of the same type within a molecule. It provides information about scalar couplings between adjacent nuclei, which helps determine connectivity and structural information. There are several COSY variants, each with its unique strengths and experimental parameters.
COSY90 is the standard two-dimensional (2D) COSY experiment that...
COSY90 is the standard two-dimensional (2D) COSY experiment that...
305
01:062D NMR: Homonuclear Correlation Spectroscopy (COSY)
1.4K
Homonuclear correlation spectroscopy, or COSY, is a 2-dimensional NMR technique that provides information about coupled protons. Typically, the geminal and vicinal coupling are observed. For example, consider the COSY spectrum of ethyl acetate, where its 1D proton NMR spectrum is plotted along the vertical and horizontal axes with their corresponding chemical shift scale. Three spots on the diagonal corresponding to the three peaks in the 1D proton spectrum are called diagonal peaks. The COSY...
1.4K
01:02¹H NMR: Interpreting Distorted and Overlapping Signals
1.1K
Spin systems where the difference in chemical shifts of the coupled nuclei is greater than ten times J are called first-order spin systems. These nuclei are weakly coupled, and their chemical shifts and coupling constant can generally be estimated from the well-separated signals in the spectrum.
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are...
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are...
1.1K