NMR methodology

Pulse sequence development

NMR spectroscopy is ideally suited to investigate Intrinsically disordered proteins (IDPs) or disordered protein regions (IDRs). As a technique, it is very sensitive to changes in molecular dynamics, and the inherent disorder incurs favourable spectroscopic properties that allow the design of very informative experiments. However, IDPs also present unique challenges to NMR spectroscopy. Compared to their globular counterparts, IDPs display significantly lower chemical shift dispersion, leading – even for small proteins – to very crowded NMR spectra, impairing extraction of valuable structural information. Moreover, the fast interconversion between conformers makes data interpretation not straightforward. We are interested in developing new approaches that harvest a maximum of spectral information that can be translated into structural information, such as residual dipolar couplings (RDCs). Central to this is the use of ‘pure shift’ techniques, whereby the signals are homodecoupled so that multiplets in the 1H spectrum of typically 10-50 Hz wide reduce to singlets of only a few Hz wide, improving resolution by an order of magnitude. Such methods have proven very valuable for small molecules, including the measurement of 1H-1H scalar couplings and RDCs (Sinnaeve et al. Angew. Chem. Int. Ed. 2015; Anal. Chem. 2018; Angew. Chem. Int. Ed. 2020), and also hold potential for IDPs.