Abstract
Protein folding is crucial for biological activity. Proteins’ failure to fold correctly underlies various pathological processes, including amyloidosis, the aggregation of insoluble proteins (e.g., lysozymes) in organs. The exact conditions that trigger the structural transition of amyloids into β-sheet-rich aggregates are poorly understood, as is the case for the amyloidogenic self-assembly pathway. Ultrasound is routinely used to destabilize a protein’s structure and enhance amyloid growth. Here, we report on an unexpected ultrasound effect on lysozyme amyloid species at different stages of aggregation: ultrasound-induced structural perturbation gives rise to nonamyloidogenic folds. Our infrared and X-ray analyses of the chemical, mechanical, and thermal effects of sound on lysozyme’s structure found, in addition to the expected ultrasound-induced damage, evidence of irreversible disruption of the β-sheet fold of fibrillar lysozyme resulting in their structural transformation into monomers with no β-sheets. This structural transition is reflected in changes in the kinetics of protein self-assembly, namely, either prolonged nucleation or accelerated fibril growth. Using solution X-ray scattering, we determined the structure, the mass fraction of lysozyme monomer, and the morphology of its filamentous assemblies formed under different sound parameters. A nanomechanical analysis of ultrasound-modified protein assemblies revealed a correlation between the β-sheet content and elastic modulus of the protein material. Suppressing one of the ultrasound-derived effects allowed us to control the structural transformations of lysozyme. Overall, our comprehensive investigation establishes the boundary conditions under which ultrasound damages protein structure and fold. This knowledge can be utilized to impose medically desirable structural modifications on amyloid β-sheet-rich proteins.
Original language | English |
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Article number | e2212849120 |
Journal | Proceedings of the National Academy of Sciences of the United States of America |
Volume | 120 |
Issue number | 3 |
DOIs | |
State | Published - 17 Jan 2023 |
Externally published | Yes |
Bibliographical note
Publisher Copyright:Copyright © 2023 the Author(s).
Funding
ACKNOWLEDGMENTS. U.S. acknowledges financial support from the Nella and Leon Benoziyo Center for Neurological Diseases.In addition,U.S.thanks the Perlman family for funding the Shimanovich Lab at the Weizmann Institute of Science: “This research was made possible in part by the historical generosity of the Harold Perlman Family.” The research work is also supported by a research grant from the Anita James Rosen Foundation and SAERI Foundation. We would like to acknowledge partial support from the GMJ Schmidt Minerva Centre of Supramolecular Architectures at the Weizmann Institute.A.K.thanks Dr. T.O.Mason for the scientific discussion. We thank the European Synchrotron Radiation Facility for providing us with access to their synchrotron radiation facilities, and we thank T. Narayanan and L.Matthews for their assistance in using the SAXS setup at beamline ID02.We acknowledge Sidney R.Cohen for his contribution to the mechanical characterization of lysozyme species, Daniel Khaykelson for the assistance in SAXS measurements, and Natalie Page for English editing.Supported by a research grant from the Estate of Betty Weneser. U.S. acknowledges financial support from the Nella and Leon Benoziyo Center for Neurological Diseases. In addition, U.S. thanks the Perlman family for funding the Shimanovich Lab at the Weizmann Institute of Science: “This research was made possible in part by the historical generosity of the Harold Perlman Family.” The research work is also supported by a research grant from the Anita James Rosen Foundation and SAERI Foundation. We would like to acknowledge partial support from the GMJ Schmidt Minerva Centre of Supramolecular Architectures at the Weizmann Institute. A.K. thanks Dr. T.O. Mason for the scientific discussion. We thank the European Synchrotron Radiation Facility for providing us with access to their synchrotron radiation facilities, and we thank T. Narayanan and L. Matthews for their assistance in using the SAXS setup at beamline ID02. We acknowledge Sidney R. Cohen for his contribution to the mechanical characterization of lysozyme species, Daniel Khaykelson for the assistance in SAXS measurements, and Natalie Page for English editing. Supported by a research grant from the Estate of Betty Weneser.
Funders | Funder number |
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Anita James Rosen Foundation | |
Estate of Betty Weneser | |
Perlman family for funding the Shimanovich Lab | |
SAERI Foundation | |
U.S.thanks the Perlman family for funding the Shimanovich Lab | |
European Synchrotron Radiation Facility | |
Weizmann Institute of Science |
Keywords
- amyloid
- beta-sheet conformation
- cavitation
- fibrillar protein self-assembly
- ultrasound