NMR-profiles of protein solutions

Authors

  • Bill Pedrini,

    1. Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA
    2. Institute of Molecular Biology and Biophysics, ETH Zürich, Zürich, Switzerland
    Current affiliation:
    1. Present address: SwissFEL Project, Paul Scherrer Institute, 5232 Villigen PSI, Switzerland and Biswaranjan Mohanty Monash University, Parkville Campus, Parkville, Australia
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    • Bill Pedrini and Pedro Serrano contributed equally to this work

  • Pedro Serrano,

    1. Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA
    2. Joint Center for Structural Genomics, The Scripps Research Institute, La Jolla, CA
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  • Biswaranjan Mohanty,

    1. Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA
    2. Joint Center for Structural Genomics, The Scripps Research Institute, La Jolla, CA
    3. Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA
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  • Michael Geralt,

    1. Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA
    2. Joint Center for Structural Genomics, The Scripps Research Institute, La Jolla, CA
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  • Kurt Wüthrich

    Corresponding author
    1. Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA
    2. Institute of Molecular Biology and Biophysics, ETH Zürich, Zürich, Switzerland
    3. Joint Center for Structural Genomics, The Scripps Research Institute, La Jolla, CA
    4. Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA
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  • This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

ABSTRACT

NMR-Profiles are quantitative one-dimensional (1D) presentations of 2D [15N,1H]-correlation spectra used to monitor the quality of protein solutions prior to and during NMR structure determinations and functional studies. In our current use in structural genomics projects, an NMR-Profile is recorded at the outset of a structure determination, using a uniformly 15N-labeled microscale sample of the protein. We thus assess the extent to which polypeptide backbone resonance assignments can be achieved with given NMR techniques, for example, conventional triple resonance experiments or APSY-NMR. With the availability of sequence-specific polypeptide backbone resonance assignments in the course of the structure determination, an “Assigned NMR-Profile” is generated, which visualizes the variation of the 15N–1H correlation cross peak intensities along the sequence and thus maps the sequence locations of polypeptide segments for which the NMR line shapes are affected by conformational exchange or other processes. The Assigned NMR-Profile provides a guiding reference during later stages of the structure determination, and is of special interest for monitoring the protein during functional studies, where dynamic features may be modulated during physiological processes. © 2013 Wiley Periodicals, Inc. Biopolymers 99: 825–831, 2013.

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