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Backbone assignment of perdeuterated proteins using long-range H/C-dipolar transfers

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Abstract

For micro-crystalline proteins, solid-state nuclear magnetic resonance spectroscopy of perdeuterated samples can provide spectra of unprecedented quality. Apart from allowing to detect sparsely introduced protons and thereby increasing the effective resolution for a series of sophisticated techniques, deuteration can provide extraordinary coherence lifetimes—obtainable for all involved nuclei virtually without decoupling and enabling the use of scalar magnetization transfers. Unfortunately, for fibrillar or membrane-embedded proteins, significantly shorter transverse relaxation times have been encountered as compared to micro-crystalline proteins despite an identical sample preparation, calling for alternative strategies for resonance assignment. In this work we propose an approach towards sequential assignment of perdeuterated proteins based on long-range 1H/13C Cross Polarization transfers. This strategy gives rise to H/N-separated correlations involving Cα, Cβ, and CO chemical shifts of both, intra- and interresidual contacts, and thus connecting adjacent residues independent of transverse relaxation times.

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Acknowledgments

I am grateful to Prof. Bernd Reif for the generous gift of the protein material. Dr. James Hook is kindly acknowledged for his support to the project. This research was financed by the Analytical Centre, UNSW.

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  1. Rasmus Linser

    Present address: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Av., Boston, MA, 02115, USA

Authors and Affiliations

  1. Analytical Centre, University of New South Wales, Sydney, NSW, 2052, Australia

    Rasmus Linser

  2. School of Chemistry, University of New South Wales, Sydney, NSW, 2052, Australia

    Rasmus Linser

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  1. Rasmus Linser
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Correspondence to Rasmus Linser.

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Linser, R. Backbone assignment of perdeuterated proteins using long-range H/C-dipolar transfers. J Biomol NMR 52, 151–158 (2012). https://doi.org/10.1007/s10858-011-9593-2

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