Journal of Biomolecular NMR

, Volume 69, Issue 4, pp 229–235 | Cite as

Label-free NMR-based dissociation kinetics determination

  • Pablo Trigo-Mouriño
  • Christian Griesinger
  • Donghan LeeEmail author


Understanding the dissociation of molecules is the basis to modulate interactions of biomedical interest. Optimizing drugs for dissociation rates is found to be important for their efficacy, selectivity, and safety. Here, we show an application of the high-power relaxation dispersion (RD) method to the determination of the dissociation rates of weak binding ligands from receptors. The experiment probes proton RD on the ligand and, therefore, avoids the need for any isotopic labeling. The large ligand excess eases the detection significantly. Importantly, the use of large spin-lock fields allows the detection of faster dissociation rates than other relaxation approaches. Moreover, this experimental approach allows to access directly the off-rate of the binding process without the need for analyzing a series of samples with increasing ligand saturation. The validity of the method is shown with small molecule interactions using two macromolecules, bovine serum albumin and tubulin heterodimers.


NMR Ligand binding Relaxation dispersion 



This work was supported by funds from the James Graham Brown Foundation, the National Center for Research Resources CoBRE (1P30GM106396), the Max Planck Society, and the EU (ERC Grant Agreement Number 233227 to C.G.). P.T.M. acknowledges the Humboldt Foundation for a postdoctoral research fellowship.

Supplementary material

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Supplementary material 1 (DOCX 1194 KB)


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2017

Authors and Affiliations

  • Pablo Trigo-Mouriño
    • 1
  • Christian Griesinger
    • 1
  • Donghan Lee
    • 2
    Email author
  1. 1.Department of NMR-Based Structural BiologyMax-Planck Institute for Biophysical ChemistryGöttingenGermany
  2. 2.James Graham Brown Cancer CenterUniversity of LouisvilleLouisvilleUSA

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