Electron Microscopy Identifies Protein Conformational Substates in Liquid Environment.

Authors

Xiaozhe Dong, Hong Zhang, Zhi Qu, Sicheng Zhao, Jia-Ye Li, Yihao Niu, Xiaoyun Pang, Qian Huang, Ye-Jin Kim, Oh-Hoon Kwon, Fei Sun, Bin Xia, Yi-Qin Gao, Steve Granick, Huan Wang

Published in

ACS nano. Volume 20. Issue 44. Pages 29983-29997. Nov 10, 2026.

Abstract

Protein functions are closely linked to their dynamic behaviors, particularly involving low-abundance transient states. Quantifying these interconversions remains experimentally challenging, especially for proteins containing intrinsically disordered regions (IDRs). In this study, we utilize low-dose electron microscopy using graphene liquid cells to image and quantify interconversions between protein conformational states under optimized electron beam conditions and confinement effects. Applied to four proteins in solution: CRISPR-associated protein (Cas9), DNA binding protein (GapR), restriction enzyme (EcoRI), and an RNA binding protein with IDRs (FUS), we visualized low-abundance transient structures distinct from crystalline forms and elucidated transition pathways which are validated by cryo-electron microscopy, revealing differences in their energy landscapes through single-molecule imaging. We analyzed domain-scale rearrangements at ∼1-2 nm effective conformational resolution, cross-validated against independent computational and experimental methods. Shifts in state equilibria and conformational adaptations were observed when proteins are mutated or during biomolecular interactions, such as target searching, binding, and cleavage with nucleic acids. Notably, changes in the energy landscape were observed when proteins with IDRs and RNA were within close proximity but not in apparent contact, suggesting that conformational redistribution may precede complete binding. This study shows how to characterize protein conformations and their time-dependent changes using instrumentation that is widely available at research universities but not yet used for these purposes.

PMID:
43007784
Bibliographic data and abstract were imported from PubMed on 11 Nov 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.