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Global Fold Determination of Large Proteins using Site-Directed Spin Labeling

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Protein NMR for the Millennium

Part of the book series: Biological Magnetic Resonance ((BIMR,volume 20))

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Abstract

Protein structures up to 25 kDa may be obtained using distance and torsion angle restraints provided by the nuclear Overhauser effect (NOE) and J couplings respectively. Alternatively, distances may be obtained between a paramagnetic center and a nucleus through the electron-nuclear dipolar interaction. A paramagnetic center may be introduced into an intrinsically diamagnetic protein by covalently attaching a nitroxide spin label to cysteine. This technique, referred to as site-directed spin labeling (SDSL), has been employed extensively in electron-paramagnetic resonance. We review applications of SDSL to obtain global folds of proteins and macromolecular complexes within the frame work of high-resolution NMR. Issues pertaining to protein engineering, interpretation of paramagnetic broadening and combination of paramagnetic broadening with limited NOE data will be discussed. Paramagnetic broadening effects (PBEs) in conjunction with limited NOE data and/or residual dipolar couplings and TROSY triple resonance methodology promise to extend the size limit of proteins amenable to structural studies by NMR

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Author information

Authors and Affiliations

  1. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, 02115

    John L. Battiste, John D. Gross & Gerhard Wagner

Authors
  1. John L. Battiste
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  2. John D. Gross
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  3. Gerhard Wagner
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Editor information

Editors and Affiliations

  1. University of Alabama at Birmingham, Birmingham, Alabama

    N. Rama Krishna

  2. University of Denver, Denver, Colorado

    Lawrence J. Berliner

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© 2002 Kluwer Academic Publishers

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Battiste, J.L., Gross, J.D., Wagner, G. (2002). Global Fold Determination of Large Proteins using Site-Directed Spin Labeling. In: Krishna, N.R., Berliner, L.J. (eds) Protein NMR for the Millennium. Biological Magnetic Resonance, vol 20. Springer, Boston, MA. https://doi.org/10.1007/0-306-47936-2_4

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  • DOI: https://doi.org/10.1007/0-306-47936-2_4

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-0-306-47448-4

  • Online ISBN: 978-0-306-47936-6

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