https://3demmethods.i2pc.es/index.php?action=history&feed=atom&title=2024Suder_Workflow 2026-05-24T22:00:59Z Revision history for this page on the wiki MediaWiki 1.44.2 https://3demmethods.i2pc.es/index.php?title=2024Suder_Workflow&diff=4661&oldid=prev 2024-08-09T06:43:12Z

<p>Created page with &quot;== Citation == Suder, Diana S. / Gonen, Shane. Mitigating the Blurring Effect of CryoEM Averaging on a Flexible and Highly Symmetric Protein Complex through Sub-Particle Reconstruction. 2024. Intl. J. Molecular Sciences, Vol. 25, No. 11, p. 5665 == Abstract == Many macromolecules are inherently flexible as a feature of their structure and function. During single-particle CryoEM processing, flexible protein regions can be detrimental to highresolution reconstruction a...&quot;</p> <p><b>New page</b></p><div>== Citation ==<br /> <br /> Suder, Diana S. / Gonen, Shane. Mitigating the Blurring Effect of CryoEM Averaging on a Flexible and Highly Symmetric Protein Complex through Sub-Particle Reconstruction. 2024. <br /> Intl. J. Molecular Sciences, Vol. 25, No. 11, p. 5665<br /> <br /> == Abstract ==<br /> <br /> Many macromolecules are inherently flexible as a feature of their structure and function.<br /> During single-particle CryoEM processing, flexible protein regions can be detrimental to highresolution<br /> reconstruction as signals from thousands of particles are averaged together. This “blurring”<br /> effect can be difficult to overcome and is possibly more pronounced when averaging highly symmetric<br /> complexes. Approaches to mitigating flexibility during CryoEM processing are becoming increasingly<br /> critical as the technique advances and is applied to more dynamic proteins and complexes. Here,<br /> we detail the use of sub-particle averaging and signal subtraction techniques to precisely target and<br /> resolve flexible DARPin protein attachments on a designed tetrahedrally symmetric protein scaffold<br /> called DARP14. Particles are first aligned as full complexes, and then the symmetry is reduced by<br /> alignment and focused refinement of the constituent subunits. The final reconstructions we obtained<br /> were vastly improved over the fully symmetric reconstructions, with observable secondary structure<br /> and side-chain placement. Additionally, we were also able to reconstruct the core region of the<br /> scaffold to 2.7 Å. The data processing protocol outlined here is applicable to other dynamic and<br /> symmetric protein complexes, and our improved maps could allow for new structure-guided variant<br /> designs of DARP14.<br /> <br /> == Keywords ==<br /> <br /> == Links ==<br /> <br /> https://www.mdpi.com/1422-0067/25/11/5665<br /> <br /> == Related software ==<br /> <br /> == Related methods ==<br /> <br /> == Comments ==</div>

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