Summary
A time-shared [15N, 13C] half-filter technique is presented, which can be used to study proton-proton NOEs between biomolecules. The filter is demonstrated in a 2D [15N, 13C] double filtered NOESY experiment of a dimeric Mnt repressor mutant consisting of completely [15N, 13C] labeled monomer and unlabeled monomer. The benefit of this combined [15N, 13C] half-filter is that a single NMR experiment can be designed that yields all NOE interactions between labeled and unlabeled protons ((13C, 14N/12C), (15N, 14N/12C), (12C, 15N/13C) and (14N, 15N/13C)) in the protein, where conventional half-filters would require at least three separate NMR experiments to obtain these NOEs. The intermonomer NOEs of the Mnt mutant confirmed the secondary structure of the DNA-binding domain as an antiparallel β ribbon, formed from an N-terminal segment contributed by each monomer. Moreover, several intersubunit NOEs were characterized in the C-terminal part of the Mnt mutant for which no structural data is available yet.
References
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Burgering, M., Boelens, R. & Kaptein, R. Observation of intersubunit NOEs in a dimeric P22 Mnt repressor mutant by a time-shared [15N, 13C] double half-filter technique. J Biomol NMR 3, 709–714 (1993). https://doi.org/10.1007/BF00198373
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DOI: https://doi.org/10.1007/BF00198373