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Issue 15, 2021
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Microchip-based structure determination of low-molecular weight proteins using cryo-electron microscopy

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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.

Graphical abstract: Microchip-based structure determination of low-molecular weight proteins using cryo-electron microscopy

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Article information


Submitted
19 Jan 2021
Accepted
21 Mar 2021
First published
01 Apr 2021

Nanoscale, 2021,13, 7285-7293
Article type
Paper

Microchip-based structure determination of low-molecular weight proteins using cryo-electron microscopy

M. A. Casasanta, G. M. Jonaid, L. Kaylor, W. Y. Luqiu, M. J. Solares, M. L. Schroen, W. J. Dearnaley, J. Wilson, M. J. Dukes and D. F. Kelly, Nanoscale, 2021, 13, 7285
DOI: 10.1039/D1NR00388G

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