Sparse Labelling for Protein NMR
Sparse labelling is a strategy in protein NMR spectroscopy where only a subset of atoms (typically isotopes like 13C or 15N) are selectively introduced into the protein, rather than uniformly labelling all possible sites.
The aim is to reduce spectral crowding and simplify analysis of large or complex proteins, especially in cases where full uniform labelling leads to overlapping signals.
It is standard practice to enrich proteins to high levels of 13C enrichment in-order to make spectral assignments and to ultimately determine structure. However, a high level of uniform 13C enrichment creates scalar and dipolar coupling between the directly bonded 13C nuclei, which for dipolar-based sequences used in solid state NMR, creates large signals which often obscures low-intensity, information-rich signals produced from long range coupling. For scalar-based sequences used in solution state NMR, the presence of directly bonded 13C nuclei will degrade spectral resolution and provides an unwanted dipolar relaxation mechanism for the alpha carbon.
Reduction of 13C-13C dipolar coupling is made possible by sparse 13C labelling. 13C sparse labelling is where the expressed protein does not contain adjacent 13C nuclei. This can be accomplished to a large degree by using selectively 13C labelled carbon sources for the protein expression. There are several different substrates available for 13C sparse labelling, including glucose, glycerol and pyruvate.
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