Metskas, Lauren Ann and Wilfong, Rosalie and Jensen, Grant J. (2022) Subtomogram averaging for biophysical analysis and supramolecular context. Journal of Structural Biology: X, 6 . Art. No. 100076. ISSN 2590-1524. PMCID PMC9596874. doi:10.1016/j.yjsbx.2022.100076. https://resolver.caltech.edu/CaltechAUTHORS:20221114-805533100.17
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Abstract
Recent advances in hardware, software and computing power have led to increasingly ambitious applications of cryo-electron tomography and subtomogram averaging. It is now possible to reveal both structures and biophysical relationships like protein binding partners and small molecule occupancy in these experiments. However, some data processing choices require the user to prioritize structure or biophysical context. Here, we present a modified subtomogram averaging approach that preserves both capabilities. By increasing the accuracy of particle-picking, performing alignment and averaging on all subtomograms, and decreasing reliance on symmetry and tight masks, the usability of tomography and subtomogram averaging data for biophysical analyses is greatly increased without negatively impacting structural refinements.
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| Additional Information: | We thank S. Chen and A. Malyutin for assistance with the tomography data collection script, and A. Burt for pseudocode to transition between Dynamo and Relion software packages. This work was supported by a Ruth L. Kirschstein NRSA Individual Postdoctoral Fellowship F32 1F32GM135994-01 to LAM, NIAID New Innovators Award 1DP2AI164293-01 to LAM, and NIH R01 AI127401 to GJJ. | |||||||||
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| PubMed Central ID: | PMC9596874 | |||||||||
| DOI: | 10.1016/j.yjsbx.2022.100076 | |||||||||
| Record Number: | CaltechAUTHORS:20221114-805533100.17 | |||||||||
| Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20221114-805533100.17 | |||||||||
| Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | |||||||||
| ID Code: | 117862 | |||||||||
| Collection: | CaltechAUTHORS | |||||||||
| Deposited By: | Research Services Depository | |||||||||
| Deposited On: | 29 Nov 2022 18:35 | |||||||||
| Last Modified: | 29 Nov 2022 18:35 |
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