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Probing Arginine Side-Chains and Their Dynamics with Carbon-Detected NMR Spectroscopy: Application to the 42 kDa Human Histone Deacetylase 8 at High pH

Authors

  • Dr. Nicolas D. Werbeck,

    1. Institute of Structural and Molecular Biology, University College London, Gower Street, WC1E 6BT, London (UK)
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  • Dr. John Kirkpatrick,

    1. Institute of Structural and Molecular Biology, University College London, Gower Street, WC1E 6BT, London (UK)
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  • Dr. D. Flemming Hansen

    Corresponding author
    1. Institute of Structural and Molecular Biology, University College London, Gower Street, WC1E 6BT, London (UK)
    • Institute of Structural and Molecular Biology, University College London, Gower Street, WC1E 6BT, London (UK)

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  • Dr. S. Djordjevic is acknowledged for providing TEV-protease. Expression constructs of T4 lysozyme mutants were kindly provided by Prof. L. E. Kay and Prof. F. W. Dahlquist. We acknowledge the Medical Research Council and the National Institute for Medical Research for access to the 800 MHz spectrometer. N.D.W. acknowledges FEBS for a long-term postdoctoral fellowship. This research is supported by the Biotechnology and Biological Sciences Research Council (BBSRC). D.F.H. is a BBSRC David Phillips Fellow.

Abstract

original image

Arginin-Seitenketten kommt durch ihren hohen pKa-Wert mit einhergehender positiver Ladung eine besondere Bedeutung zu. Eine NMR-Methode basierend auf Kohlenstoff-detektierten 13Cζ-15Nε-Korrelationsspektren wird vorgestellt, welche die Detektion von Arginin-Seitenketten im neutralen Bereich bis hin zu hohem pH ermöglicht. Die Methode wird an der humanen Histon-Deacetylase 8 demonstriert.

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