Suppression of sampling artefacts in high-resolution four-dimensional NMR spectra using Signal Separation Algorithm

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Suppression of sampling artefacts in high-resolution four-dimensional NMR spectra using Signal Separation Algorithm


Publication year: 2011
Source: Journal of Magnetic Resonance, Available online 20 October 2011

Jan*Stanek, Rafal*Augustyniak, Wiktor*Ko?mi?ski

The development of non-uniform sampling (NUS) strategies permits to obtain high-dimensional spectra with increased resolution in significantly reduced experimental time. We extended a previously proposed signal separation algorithm (SSA) to process sparse four-dimensional NMR data. It is employed for two experiments carried out for a partially unstructured 114-residue construct of chicken Engrailed 2 protein, namely 4D HCCH-TOCSY and 4D C,*N-edited NOESY. The SSA allowed us to obtain high-quality spectra using only as little as 0.16% of the available samples, with low sampling artefacts approaching the thermal noise level in most spectral regions. It is demonstrated that NUS 4D HCCH-TOCSY is dominated by sampling noise and requires efficient artefact suppression. On the other hand, 4D C,*N-edited NOESY is a particularly attractive experiment for NUS, as the absence of diagonal peaks renders the problem of artefacts less critical. We also present a transverse-relaxation optimized sequence for HMQC that is especially designed for longer evolution periods in the indirectly detected proton dimension in high-dimensional pulse sequences. In conjunction with novel sampling strategies and efficient processing methods, this improvement enabled us to obtain unique structural information about aliphatic-amide contacts.

Graphical abstract



Highlights

? A four-dimensional version of Signal Separation Algorithm has been presented. ? 4D HCCH-TOCSY and 4D C,N-edited NOESY were performed on a 13.4 kDa protein. ? Up to 90% sampling noise reduction was achieved in 4D HCCH-TOCSY. ? Sampling density as low as 0.16% were used. ? A transverse relaxation optimization of HMQC increases sensitivity up to 18%.



Source: Journal of Magnetic Resonance