5D solid-state NMR spectroscopy for facilitated resonance assignment

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dc.contributor.authorKlein, Alexander
dc.contributor.authorVasa, Suresh K.
dc.contributor.authorLinser, Rasmus
dc.date.accessioned2025-05-27T09:04:01Z
dc.date.available2025-05-27T09:04:01Z
dc.date.issued2023-11-09
dc.description.abstract1H-detected solid-state NMR spectroscopy has been becoming increasingly popular for the characterization of protein structure, dynamics, and function. Recently, we showed that higher-dimensionality solid-state NMR spectroscopy can aid resonance assignments in large micro-crystalline protein targets to combat ambiguity (Klein et al., Proc. Natl. Acad. Sci. U.S.A. 2022). However, assignments represent both, a time-limiting factor and one of the major practical disadvantages within solid-state NMR studies compared to other structural-biology techniques from a very general perspective. Here, we show that 5D solid-state NMR spectroscopy is not only justified for high-molecular-weight targets but will also be a realistic and practicable method to streamline resonance assignment in small to medium-sized protein targets, which such methodology might not have been expected to be of advantage for. Using a combination of non-uniform sampling and the signal separating algorithm for spectral reconstruction on a deuterated and proton back-exchanged micro-crystalline protein at fast magic-angle spinning, direct amide-to-amide correlations in five dimensions are obtained with competitive sensitivity compatible with common hardware and measurement time commitments. The self-sufficient backbone walks enable efficient assignment with very high confidence and can be combined with higher-dimensionality sidechain-to-backbone correlations from protonated preparations into minimal sets of experiments to be acquired for simultaneous backbone and sidechain assignment. The strategies present themselves as potent alternatives for efficient assignment compared to the traditional assignment approaches in 3D, avoiding user misassignments derived from ambiguity or loss of overview and facilitating automation. This will ease future access to NMR-based characterization for the typical solid-state NMR targets at fast MAS.en
dc.identifier.urihttp://hdl.handle.net/2003/43708
dc.identifier.urihttp://dx.doi.org/10.17877/DE290R-25482
dc.language.isoen
dc.relation.ispartofseriesJournal of biomolecular NMR; 77(5/6)
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectSolid-state NMRen
dc.subjectProton detectionen
dc.subjectFast magic-angle spinningen
dc.subjectHigher dimensionalityen
dc.subject5Den
dc.subjectResonance assignmenten
dc.subjectMinimal set of experimentsen
dc.subjectNon-uniform samplingen
dc.subject.ddc540
dc.title5D solid-state NMR spectroscopy for facilitated resonance assignmenten
dc.typeText
dc.type.publicationtypeArticle
dcterms.accessRightsopen access
eldorado.secondarypublicationtrue
eldorado.secondarypublication.primarycitationKlein, A., Vasa, S.K. & Linser, R. 5D solid-state NMR spectroscopy for facilitated resonance assignment. J Biomol NMR 77, 229–245 (2023). https://doi.org/10.1007/s10858-023-00424-5
eldorado.secondarypublication.primaryidentifierhttps://doi.org/10.1007/s10858-023-00424-5

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