Uncovering the Conformational Distribution of a Small Protein with Nanoparticle-Aided Cryo-Electron Microscopy Sampling
- 12 July 2021
- journal article
- research article
- Published by American Chemical Society (ACS) in The Journal of Physical Chemistry Letters
- Vol. 12 (28), 6565-6573
- https://doi.org/10.1021/acs.jpclett.1c01277
Abstract
Here, we introduce the nanoparticle-aided cryo-electron microscopy sampling (NACS) method to access the conformational distribution of a protein molecule. Two nanogold particles are labeled at two target sites, and the interparticle distance is measured as a structural parameter via cryo-electron microscopy (cryo-EM). The key aspect of NACS is that the projected distance information instead of the global conformational information is extracted from each protein molecule. This is possible because the contrast provided by the nanogold particles is strong enough to provide the projected distance, while the protein itself is invisible due to its low contrast. We successfully demonstrate that various protein conformations, even for small or disordered proteins, which generally cannot be accessed via cryo-EM, can be captured. The demonstrated method with the potential to directly observe the conformational distribution of such systems may open up new possibilities in studying their dynamics at a single-molecule level.Funding Information
- Samsung Science and Technology Foundation (SSTF-BA1802-05)
- National Research Foundation of Korea (2018M3D1A1058813)
- Institute for Basic Science (IBS-R004)
This publication has 47 references indexed in Scilit:
- 3D Motion of DNA-Au Nanoconjugates in Graphene Liquid Cell Electron MicroscopyNano Letters, 2013
- Manipulating Protein Conformations by Single-Molecule AFM-FRET NanoscopyACS Nano, 2012
- Site-Specific Biomolecule Labeling with Gold ClustersMethods in Enzymology, 2010
- Site-directed nanoparticle labeling of cytochrome cProceedings of the National Academy of Sciences of the United States of America, 2009
- Biophysical characterization of intrinsically disordered proteinsCurrent Opinion in Structural Biology, 2009
- Tight Regulation of Unstructured Proteins: From Transcript Synthesis to Protein DegradationScience, 2008
- Protein folding studied by single-molecule FRETCurrent Opinion in Structural Biology, 2008
- Intrinsic Disorder in Transcription FactorsBiochemistry, 2006
- Intrinsically unstructured proteins and their functionsNature Reviews Molecular Cell Biology, 2005
- Intrinsic Disorder in Cell-signaling and Cancer-associated ProteinsJournal of Molecular Biology, 2002