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Identification of macromolecular complexes in cryoelectron tomograms of phantom cells
Open accessAbstract
Electron tomograms of intact frozen-hydrated cells are essentially three-dimensional images of the entire proteome of the cell, and they depict the whole network of macromolecular interactions. However, this information is not easily accessible because of the poor signal-to-noise ratio of the tomograms and the crowded nature of the cytoplasm. Here, we describe a template matching algorithm that is capable of detecting and identifying macromolecules in tomographic volumes in a fully automated manner. The algorithm is based on nonlinear cross correlation and incorporates elements of multivariate statistical analysis. Phantom cells, i.e., lipid vesicles filled with macromolecules, provide a realistic experimental scenario for an assessment of the fidelity of this approach. At the current resolution of ≈4 nm, macromolecules in the size range of 0.5–1 MDa can be identified with good fidelity.
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Citations
Cites
Macromolecular crowding: obvious but underappreciated
Ellis, R. J. (2001) · Trends in Biochemical Sciences · doi:10.1016/s0968-0004(01)01938-7 · Cited by 2,309
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Three-dimensional architecture of murine rod outer segments determined by cryoelectron tomography
Nickell, S., Park, P. S.-H., Baumeister, W., & Palczewski, K. (2007) · The Journal of Cell Biology · doi:10.1083/jcb.200612010 · Cited by 209
Self-citationIntegrating diverse data for structure determination of macromolecular assemblies
Alber, F., Förster, F., Korkin, D., Topf, M., & Sali, A. (2008) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.77.060407.135530 · Cited by 206
Self-citationVisual proteomics of the human pathogen Leptospira interrogans
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Metadata
- Authors
- Frangakis, A. S., Böhm, J., Förster, F., Nickell, S., Nicastro, D., Typke, D., Hegerl, R., & Baumeister, W.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2002-10-21
- Volume
- 99
- Issue
- 22
- Pages
- 14153-14158
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2003294049
- Cited by
- 267
- References
- 1
- FWCI
- 7.51
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Frangakis, A. S., Böhm, J., Förster, F., Nickell, S., Nicastro, D., Typke, D., Hegerl, R., & Baumeister, W. (2002). Identification of macromolecular complexes in cryoelectron tomograms of phantom cells. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.172520299
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