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Article Dans Une Revue Journal of Biomolecular NMR Année : 2020

Spectral editing of intra- and inter-chain methyl–methyl NOEs in protein complexes

Résumé

Specific isotopic labeling of methyl groups in a perdeuterated protein background enables the detection of long range NOEs in proteins or high molecular weight complexes. We introduce here an approach, combining an optimized isotopic labeling scheme with a specifically tailored NMR pulse sequence, to distinguish between intramolecular and intermolecular NOE connectivities. In hetero-oligomeric complexes, this strategy enables sign encoding of intra-subunit and inter-subunit NOEs. For homo-oligomeric assemblies, our strategy allows the specific detection of intra-chain NOEs in high resolution 3D NOESY spectra. The general principles, possibilities and limitations of this approach are presented. Applications of this approach for the detection of intermolecular NOEs in a hetero-hexamer, and the assignment of methyl 1H and 13C resonances in a homo-tetrameric protein complex are shown.
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Dates et versions

hal-02504779 , version 1 (21-06-2021)

Identifiants

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Ricarda Törner, Rida Awad, Pierre Gans, Bernhard Brutscher, Jérôme Boisbouvier. Spectral editing of intra- and inter-chain methyl–methyl NOEs in protein complexes. Journal of Biomolecular NMR, 2020, 74 (1), pp.83-94. ⟨10.1007/s10858-019-00293-x⟩. ⟨hal-02504779⟩
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