Quantifying the heterogeneity of macromolecular machines by mass photometry
Name
s41467-020-15642-w.pdf
Description
Published version
Size
2.08 MB
Format
Adobe PDF
Checksum (MD5)
016e7b9c6be1536d50383eee806f8cce
Author(s)
Sonn-Segev, Adar
Belacic, Katarina
Bodrug, Tatyana
Young, Gavin
VanderLinden, Ryan T
Schulman, Brenda A
Schimpf, Johannes
Friedrich, Thorsten
Dip, Phat Vinh
Schwartz, Thomas U
Date Issued
2020
Journal
Nature Communications
Publisher
Springer Science and Business Media LLC
Version
Final published version
Abstract
© 2020, The Author(s). Sample purity is central to in vitro studies of protein function and regulation, and to the efficiency and success of structural studies using techniques such as x-ray crystallography and cryo-electron microscopy (cryo-EM). Here, we show that mass photometry (MP) can accurately characterize the heterogeneity of a sample using minimal material with high resolution within a matter of minutes. To benchmark our approach, we use negative stain electron microscopy (nsEM), a popular method for EM sample screening. We include typical workflows developed for structure determination that involve multi-step purification of a multi-subunit ubiquitin ligase and chemical cross-linking steps. When assessing the integrity and stability of large molecular complexes such as the proteasome, we detect and quantify assemblies invisible to nsEM. Our results illustrate the unique advantages of MP over current methods for rapid sample characterization, prioritization and workflow optimization.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1038/S41467-020-15642-W
