Solid-State NMR Spectroscopy of Functional Amyloid from a Fungal Hydrophobin: A Well-Ordered β-Sheet Core Amidst Structural Heterogeneity

Authors

  • Vanessa K. Morris,

    1. School of Medical Sciences and School of Molecular Bioscience, University of Sydney, Sydney (Australia)
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    • These authors contributed equally to this work.

  • Rasmus Linser,

    1. School of Chemistry, University of New South Wales, Sydney (Australia)
    2. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston (USA)
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    • These authors contributed equally to this work.

  • Karyn L. Wilde,

    1. National Deuteration Facility, Australian Nuclear Science and Technology Organisation, Lucas Heights (Australia)
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  • Anthony P. Duff,

    1. National Deuteration Facility, Australian Nuclear Science and Technology Organisation, Lucas Heights (Australia)
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  • Margaret Sunde,

    Corresponding author
    1. School of Medical Sciences and School of Molecular Bioscience, University of Sydney, Sydney (Australia)
    • School of Medical Sciences and School of Molecular Bioscience, University of Sydney, Sydney (Australia)
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  • Ann H. Kwan

    Corresponding author
    1. School of Medical Sciences and School of Molecular Bioscience, University of Sydney, Sydney (Australia)
    • School of Medical Sciences and School of Molecular Bioscience, University of Sydney, Sydney (Australia)
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  • We thank Dr. J. Hook, Dr. D. Thomas, and Dr. D. Lawes from the NMR facility at University of New South Wales for spectrometer access and assistance, Dr. N. Shepherd and Dr. B. Crossett for mass spectrometry at the Sydney University Proteome Research Unit, and Dr. A. Kondyurin for assistance with contact angle measurements. The authors acknowledge the facilities, and the scientific and technical assistance, of the Australian Microscopy & Microanalysis Research Facility at the Electron Microscope Unit, University of Sydney. The HET-s spectrum was kindly provided by Dr. C. Wasmer and Prof. B. Meier. The project was funded by the Australian Research Council (LP0776672 and DP0879121) and ANSTO Bragg Institute (NDF 1668). M.S. was supported by a National Health and Medical Research Council R.D. Wright Career Development Fellowship, V.K.M. was supported by a University of Sydney Vice-Chancellor’s Research Scholarship, and R.L. was supported by an Australian Research Council Discovery Early Career Research Award.

Abstract

original image

GrEASy fibrils: Hydrophobins are fungal proteins that assemble into an amphipathic fibrillar monolayer with amyloid properties and a hydrophobic face as water-resistant as Teflon. Solid-state NMR studies on EAS hydrophobin fibrils reveal direct evidence of a partial molecular rearrangement on assembly and an ordered β-sheet-rich core in the context of a whole protein in this functional amyloid.

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