Methods to produce protein amyloid fibrils, in vitro, and in situ structure characterization, are of primary importance in biology, medicine, and pharmacology. We first demonstrated the droplet on a super-hydrophobic substrate as the reactor to produce protein amyloid fibrils with real-time monitoring of the growth process by using combined light-sheet microscopy and thermal imaging. The molecular structures were characterized by Raman spectroscopy, X-ray diffraction and X-ray scattering. We demonstrated that the convective flow induced by the temperature gradient of the sample is the main driving force in the growth of well-ordered protein fibrils. Particular attention was devoted to PHF6 peptide and full-length Tau441 protein to form amyloid fibrils. By a combined experimental with the molecular dynamics simulations, the conformational polymorphism of these amyloid fibrils were characterized. The study provided a feasible procedure to optimize the amyloid fibrils formation and characterizations of other types of proteins in future studies.
KAUST Repository Item: Exported on 2020-10-01
Acknowledged KAUST grant number(s): OCRF-2014-CRG, OCRF-2016-CRG
Acknowledgements: The authors acknowledge financial support from King Abdullah University of Science and Technology for OCRF-2014-CRG and OCRF-2016-CRG grants and from Piedmont Region through European Funds for Regional Development (“Food Digital Monitoring” project). And Istituto di Cristallografia—Consiglio Nazionale delle Ricerche (IC-CNR) would like to thank the funding from MIUR (Italian Ministry for Education, University and Re-search) in the “PON Ricerca e Competitività 2007–2013” Program: ReCaS (Azione I—Interventi di rafforzamento strutturale, PONa3_00052, Avviso 254/Ric) and PRISMA (Asse II—Sostegno ll’innovazione, PON04a2_A).