Saturation-Transfer Difference is one of the most popular techniques used with NMR to study the ligand and protein, interaction. This technique focus more on the signals emitted from the ligand molecules but not the receptor.
On this paper, the authors demonstrate how to carry out a Saturation-Transfer Difference NMR experiment in a classroom environment. The compound and molecules that they used to demonstrate this particular technique are human serum albumin, 6-D,L-methyl-tryptophan, and 7-D,L-methyl-tryptophan. These compound will be examined in term of ligand screening, dissociation constant and such.
The experiment includes several steps.
- Preparation of the Samples
- STD NMR Experimental set up
- STD-NMR Ligand Based-Screening Experiment set up
- STD Build-Up Experiment and Ligand Mapping set up
- Determination of Kd (dissociation constant)
Using the protocol attached with this paper. The students will be able to carry out this particular experiment. The degree of difficulties can also be adjusted to fit the specific type of courses and major (chemistry vs. biochemistry)
Aldino Viegas, Jo~ao Manso, Franklin L. Nobrega, and Eurico J. Cabrita (2011). Saturation-Transfer Difference (STD) NMR: A Simple and Fast Method for Ligand Screening and Characterization of Protein Binding. Jounal of Chemical Education (88): 900-994
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