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Efficient assignment of methyl resonances: Enhanced sensitivity by gradient selection in a DE-MQ–(H)CCmHt m–TOCSY experiment

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Abstract

We present a gradient selected and doubly sensitivity-enhanced DE-MQ–(H)CCmHm–TOCSY experiment for the sequence-specific assignment of methyl resonances in 13C,15N labeled proteins. The proposed experiment provides improved sensitivity and artifact suppression relative to the phase-cycled experiments. One part of the 13Cchemical shift evolution takes place under heteronuclear multiple quantum coherence, whereas the other part occurs under 13C single quantum coherence in a semi-constant time fashion. The feasibility of the experiment was assessed using 15N,13C labeled Mus musculus coactosin (16 kDa), having a rotational correlation time of 14.5 ns at 15 °C in D2O. A 16-h experiment on 600 MHz 1H yielded good quality data and enabled the assignment of 70 out of 72 methyl groups in coactosin. As well as being an improved approach for methyl resonance assignment, this experiment can also be highly valuable for the rapid assignment of methyl resonances in SAR by NMR studies.

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Authors and Affiliations

  1. NMR Laboratory, Structural Biology and Biophysics Program, Institute of Biotechnology, University of Helsinki, P.O. Box 65, Helsinki, FIN-00014, Finland

    Perttu Permi, Helena Tossavainen & Maarit Hellman

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  1. Perttu Permi
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  2. Helena Tossavainen
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  3. Maarit Hellman
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Correspondence to Perttu Permi.

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Permi, P., Tossavainen, H. & Hellman, M. Efficient assignment of methyl resonances: Enhanced sensitivity by gradient selection in a DE-MQ–(H)CCmHt m–TOCSY experiment. J Biomol NMR 30, 275–282 (2004). https://doi.org/10.1007/s10858-004-3222-2

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  • DOI: https://doi.org/10.1007/s10858-004-3222-2

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