Ligand Screening


NMRFAM is leveraging its existing inventory of metabolites to develop an NMR-based high-throughput ligand screening platform. Identification of binding interactions can lead to functional assignment of orphan proteins, identification of allosteric interactions, or establishment of drug leads. The primary screen uses 1D proton (1H) NMR spectroscopy to observe signals from the metabolite in the presence and absence of a protein or other biomacromolecule. Changes in the spectrum of the small molecule (e.g., peak broadening or loss of intensity) in the presence of the macromolecule reveal binding.

If the metabolite is chemically modified by the macromolecule, its pattern of chemical shifts will change. At left, a protein supplied by the laboratory of Dave Pagliarini (University of Wisconsin-Madison) was shown to convert ADP to AMP, a reaction that accelerated when magnesium was added.

The advantages of this NMR technique is that no labeling or biochemical assay is required and that relatively weak binding interactions can be detected.


Efficient screening is accomplished using in-house developed NMRmix software to design mixtures, each containing 4-5 compounds, with minimal peak overlap as determined by reference to 1D 1H NMR spectra in the BMRB metabolite library. NMRbot software handles automated NMR data collection. NMRscreen software (in development) analyzes the data to identify bound ligands. These technologies enable the screening of a macromolecule or complex against ~500 compounds in a single day. The amount of protein required is on the order of 3‒5 mg.

When possible, hits from this screen can be confirmed by collecting two-dimensional 1H-15N NMR data from the protein labeled with 15N. Signals perturbed upon the addition of ligand can identify its binding site (at right, protein supplied by the laboratory of Gaya Amarasinghe, Washington University, St. Louis). For larger proteins (up to 150 kDa), the approach is to label methyl groups of the protein with 13C. The methyl signals tend to be sharp (very large proteins may require a background of deuterium labeling) and can be followed by two-dimensional 1H-13C NMR.

The current library consists mainly of known physiological substrates, products, cofactors, and allosteric effectors, supplemented with some drug-like compounds and potential environmental toxins. In addition, binding of metals including calcium, cobalt, chromium, copper, iron, magnesium, manganese, molybdenum, nickel, selenium, and zinc can be evaluated by Differential Scanning Fluorimetry. The compound library will soon be expanded to more than 1000 compounds.



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Biochemistry 800 - Practical Nuclear Magnetic Resonance Theory 

Biochemistry 801 - Biochemical Applications of Nuclear Magnetic Resonance 

"The Future of NMR-Based Metabolomics, Current Opinion in Biotechnology (2017), pp. 34-40

Documents on the use of the Bruker-Axs Nanostar,SAXS instrument and analysis of SAXS data are now available.

NMRFAM-SPARKY Distribution - the popular NMR analysis program SPARKY recompiled (including updated python and Tcl/Tk) with incorporation of PINE-Sparky, enhancements to import/export to the structural analysis program CYANA, and other useful python extensions.

ADAPT-NMR Enhancer: Complete Package for Reduced Dimensionality in Protein NMR Spectroscopy

RNA-PAIRS: RNA Probabilistic Assignment of Imino Resonance Shifts

PACSY, a Relational Database Management System for Protein Structure and Chemical Shift Analysis 


Donate to NMRFAM. US tax-deductible donation can be made to NMRFAM
Please write check payable to "UW Foundation, Account 112152802"  
And mail to: 
Attn: Sarah Lynn Traver Saunders
Associate Administrative Program Specialist 
University of Wisconsin-Madison 
433 Babcock Drive 
Madison, WI 53706 
Tel: 608-265-2507 or email 


1st: Lai Bergeman 
Rm 171; Phone 262-3173

2nd: Milo Westler
Rm B160; Phone 263-9599

3rd: Paulo F. Cobra
Rm B224; Phone 265-3303

4th: Marco Tonelli
Rm B160; Phone 263-9493

5th: John Markley
Rm 171A; Phone 263-9349

We welcome your questions and feedback!

NMRFAM Established 1987