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Asor et al 2020.pdf (24.97 MB)

Rapidly Forming Early Intermediate Structures Dictate the Pathway of Capsid Assembly

submitted on 04.02.2020, 13:39 and posted on 05.02.2020, 08:37 by Roi Asor, Christopher John Schlicksup, Zhongchao Zhao, Adam Zlotnick, Uri Raviv
There are about 1030 possible intermediates on the assembly path from hepatitis B capsid protein dimers to the 120-dimer capsid. If every intermediate was tested, assembly would often get stuck in an entropic trap and essentially every capsid would follow a unique assembly path. Yet, capsids assemble rapidly with minimal trapped intermediates, a realization of the Levinthal paradox. To understand the fundamental mechanisms of capsid assembly it is critical to resolve the early stages of the reaction. We have used Time-Resolved Small Angle X-ray Scattering, which is sensitive to solute size and shape and has millisecond temporal resolution. Scattering curves were fit to a thermodynamically curated library of assembly intermediates, using the principle of maximum entropy. Maximum entropy also provides a physical rationale for the selection of species. We found that the capsid assembly pathway was exquisitely sensitive to initial assembly conditions. With the mildest conditions tested, the reaction appeared two-state from dimer to 120-dimer capsid with some dimers-of-dimers and trimers-of-dimers. In slightly more aggressive conditions, we observed transient accumulation of a decamer-of-dimers and appearance of 90-dimer capsids. In conditions where there is measurable kinetic trapping, we found that a highly diverse early intermediates accumulated within a fraction of a second and propagated into long-lived kinetically trapped states (>90-mer). In all cases, intermediates between 35 and 90 subunits did not accumulate. These results are consistent with the presence of low barrier paths that connect early and late intermediates and direct the ultimate assembly path to late intermediates where assembly can be paused.


NIH (award number 1R01AI118933), Israel Science Foundation (grant 656/17),


Email Address of Submitting Author


Institute of Chemistry, The Hebrew University of Jerusalem



ORCID For Submitting Author


Declaration of Conflict of Interest

No conflict of interest

Version Notes

First version