期刊
NATURE COMMUNICATIONS
卷 11, 期 1, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-020-15642-w
关键词
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资金
- ERC [PHOTOMASS 819593]
- NIH [T32GM008570, R37GM065930, P30CA021765, R01GM77537, R35GM128855, P30AI050410]
- NSF [DGE-1650116]
- Zvi and Ofra Meitar Magdalen Graduate Scholarship
- American Lebanese Syrian Associated Charities
- Howard Hughes Medical Institute HHMI
- Deutsche Forschungsgemeinschaft [278002225/RTG 2202, FR 1140/11-2]
- long term EMBO fellowship
- HFSP fellowship
- Boehringer Ingelheim
- Austrian Research Promotion Agency (Headquarter grant) [FFG-852936]
- ERC under the European Union's Horizon 2020 research and innovation programme GA [693949]
- HFSP [RGP0057/2018]
- Refeyn Ltd.
- UCRF
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. Mass photometry is a label-free optical approach capable of detecting, imaging and accurately measuring the mass of single biomolecules in solution. Here, the authors demonstrate the potential of mass photometry for quantitatively characterizing sample heterogeneity of purified protein complexes with implications for structural studies specifically and in vitro studies more generally.
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