Sequential acquisition of multi-dimensional heteronuclear chemical shift correlation spectra with 1H detection

Peter Bellstedt,Anika Kirschstein,Matthias Görlach,Christoph Wiedemann,Christian Herbst,Ramadurai Ramachandran,Yvonne Ihle

doi:10.1038/srep04490

Abstract

RF pulse schemes for the simultaneous acquisition of heteronuclear multi-dimensional chemical shift correlation spectra, such as {HA(CA)NH & HA(CACO)NH}, {HA(CA)NH & H(N)CAHA} and {H(N)CAHA & H(CC)NH}, that are commonly employed in the study of moderately-sized protein molecules, have been implemented using dual sequential 1H acquisitions in the direct dimension. Such an approach is not only beneficial in terms of the reduction of experimental time as compared to data collection via two separate experiments but also facilitates the unambiguous sequential linking of the backbone amino acid residues. The potential of sequential 1H data acquisition procedure in the study of RNA is also demonstrated here.

Highlights

RF pulse schemes for the simultaneous acquisition of heteronuclear multi-dimensional chemical shift correlation spectra, such as {HA(CA)NH & HA(CACO)NH}, {HA(CA)NH & H(N)CAHA} and {H(N)CAHA & H(CC)NH}, that are commonly employed in the study of moderately-sized protein molecules, have been implemented using dual sequential 1H acquisitions in the direct dimension
In protein NMR a variety of multi-dimensional heteronuclear chemical shift correlation experiments are typically used for resonance assignment and for extraction of 1H-1H distance restraints, e.g. HACANH1,2, HNCAHA3–5, HACACONH2, HNCOCAHA4,5, HCCNH1,6 and 15N-edited 1H-1H NOESY, respectively
We have reported RF pulse schemes involving dual sequential 1H acquisition with only amide proton detection and making use of dual 15N-13C mixing steps for achieving protein resonance assignment[22]. While such sequences can be adapted to the study of large 2H-labeled protein samples, here we present RF pulse schemes that were developed in the context of moderately sized proteins

Summary

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RF pulse schemes for the simultaneous acquisition of heteronuclear multi-dimensional chemical shift correlation spectra, such as {HA(CA)NH & HA(CACO)NH}, {HA(CA)NH & H(N)CAHA} and {H(N)CAHA & H(CC)NH}, that are commonly employed in the study of moderately-sized protein molecules, have been implemented using dual sequential 1H acquisitions in the direct dimension. We have reported RF pulse schemes involving dual sequential 1H acquisition with only amide proton detection and making use of dual 15N-13C mixing steps for achieving protein resonance assignment[22] While such sequences can be adapted to the study of large 2H-labeled protein samples, here we present RF pulse schemes that were developed in the context of moderately sized proteins. The efficacy of the approach is experimentally www.nature.com/scientificreports (a)

Results and discussion

Methods

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