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Microchip-based structure determination of low-molecular weight proteins using cryo-electron microscopy.
Casasanta, Michael A; Jonaid, G M; Kaylor, Liam; Luqiu, William Y; Solares, Maria J; Schroen, Mariah L; Dearnaley, William J; Wilson, Jarad; Dukes, Madeline J; Kelly, Deborah F.
  • Casasanta MA; Department of Biomedical Engineering, Pennsylvania State University, University Park, PA 16802, USA. Debkelly@psu.edu.
Nanoscale ; 13(15): 7285-7293, 2021 Apr 21.
Article in English | MEDLINE | ID: covidwho-1199322
ABSTRACT
Interest in cryo-Electron Microscopy (EM) imaging has skyrocketed in recent years due to its pristine views of macromolecules and materials. As advances in instrumentation and computing algorithms spurred this progress, there is renewed focus to address specimen-related challenges. Here we contribute a microchip-based toolkit to perform complementary structural and biochemical analysis on low-molecular weight proteins. As a model system, we used the SARS-CoV-2 nucleocapsid (N) protein (48 kDa) due to its stability and important role in therapeutic development. Cryo-EM structures of the N protein monomer revealed a flexible N-terminal "top hat" motif and a helical-rich C-terminal domain. To complement our structural findings, we engineered microchip-based immunoprecipitation assays that led to the discovery of the first antibody binding site on the N protein. The data also facilitated molecular modeling of a variety of pandemic and common cold-related coronavirus proteins. Such insights may guide future pandemic-preparedness protocols through immuno-engineering strategies to mitigate viral outbreaks.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Cryoelectron Microscopy / Coronavirus Nucleocapsid Proteins / SARS-CoV-2 Language: English Journal: Nanoscale Year: 2021 Document Type: Article Affiliation country: D1nr00388g

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Cryoelectron Microscopy / Coronavirus Nucleocapsid Proteins / SARS-CoV-2 Language: English Journal: Nanoscale Year: 2021 Document Type: Article Affiliation country: D1nr00388g