CaltechAUTHORS
Published July 7, 2023 | Version Published + Supplemental Material
Journal Article Open

Single-molecule visualization determines conformational substate ensembles in β-sheet–rich peptide fibrils

Creators

  • 1. ROR icon Harvard University
  • 2. ROR icon The Ohio State University
  • 3. ROR icon Jiangsu University
  • 4. ROR icon Peking University
  • 5. ROR icon Fujian Normal University
  • 6. ROR icon National Center for Nanoscience and Technology
  • 7. ROR icon California Institute of Technology

Abstract

An understanding of protein conformational ensembles is essential for revealing the underlying mechanisms of interpeptide recognition and association. However, experimentally resolving multiple simultaneously existing conformational substates remains challenging. Here, we report the use of scanning tunneling microscopy (STM) to analyze the conformational substate ensembles of β sheet peptides with a submolecular resolution (in-plane <2.6 Å). We observed ensembles of more than 10 conformational substates (with free energy fluctuations between several k_BTs) in peptide homoassemblies of keratin (KRT) and amyloidal peptides (−5Aβ42 and TDP-43 341–357). Furthermore, STM reveals a change in the conformational ensemble of peptide mutants, which is correlated with the macroscopic properties of peptide assemblies. Our results demonstrate that the STM-based single-molecule imaging can capture a thorough picture of the conformational substates with which to build an energetic landscape of interconformational interactions and can rapidly screen conformational ensembles, which can complement conventional characterization techniques.

Additional Information

© 2023 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). Chenxuan Wang thanks the Beijing Natural Science Foundation (2222039), the National Natural Science Foundation of China (32201243, 32201142, and 31901007), the CAMS Innovation Fund for Medical Sciences (2022-I2M-JB-003 and 2021-I2M-1-022), Haihe Laboratory of Cell Ecosystem Innovation Fund (HH22KYZX0011), the National Key Research and Development Program of China (SQ2022YFA1200125), State Key Laboratory Special Fund (2060204), Fundamental Research Funds for the Central Universities (3332021039), and Overseas Expertise Introduction Center for Discipline Innovation ("111Center") (BP0820029). X.W. thanks the funding support by the startup funds of the Ohio State University. Author contributions: Conceptualization: Chenxuan Wang, W.Z., L.Y., and X.W.; validation and investigation: W.Z., R.W., M.L., L.Y., F.Z., S. Li, Y.W., Z.L., Y.Z., and S. Liu; resources: Chenxuan Wang, W.Z., and L.Y.; data curation: Chenxuan Wang; writing—original draft: Chenxuan Wang and W.Z.; writing—review and editing: Chenxuan Wang, W.Z., L.Y., X.W., Y. Yao, S. Li, A.E.V., W.D., R.L.D., Y. Yang, and Chen Wang; visualization: R.W., M.L., W.Z., L.Y., and Chenxuan Wang; supervision and funding acquisition: Chenxuan Wang, W.Z., L.Y., and X.W. Competing interests: Chenxuan Wang, L.Y., R.W., Chen Wang, Y. Yang, W.Z., S. Li, M.L., and F.Z. are inventors on a pending Chinese patent related to this work filed by the Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, and National Center for Nanoscience and Technology (patent no. ZL 2022 1 0848779.7). The authors declare no other competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials.

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Additional details

Identifiers

PMCID
PMC10321740
Eprint ID
122508
Resolver ID
CaltechAUTHORS:20230725-705826000.2

Funding

Beijing Natural Science Foundation
2222039
National Natural Science Foundation of China
32201243
National Natural Science Foundation of China
32201142
National Natural Science Foundation of China
31901007
CAMS Innovation Fund for Medical Sciences
2022-I2M-JB-003
CAMS Innovation Fund for Medical Sciences
2021-I2M-1-022
Haihe Laboratory of Cell Ecosystem Innovation Fund
HH22KYZX0011
National Key Research and Development Program of China
SQ2022YFA1200125
State Key Laboratory Special Fund
2060204
Fundamental Research Funds for the Central Universities
3332021039
Overseas Expertise Introduction Center for Discipline Innovation
BP0820029
Ohio State University

Dates

Created
2023-08-16
Created from EPrint's datestamp field
Updated
2023-08-16
Created from EPrint's last_modified field