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Determination of the molecular weight of proteins in solution from a single small-angle X-ray scattering measurement on a relative scale
Abstract
This paper describes a new and simple method to determine the molecular weight of proteins in dilute solution, with an error smaller than ∼10%, by using the experimental data of a single small-angle X-ray scattering (SAXS) curve measured on a relative scale. This procedure does not require the measurement of SAXS intensity on an absolute scale and does not involve a comparison with another SAXS curve determined from a known standard protein. The proposed procedure can be applied to monodisperse systems of proteins in dilute solution, either in monomeric or multimeric state, and it has been successfully tested on SAXS data experimentally determined for proteins with known molecular weights. It is shown here that the molecular weights determined by this procedure deviate from the known values by less than 10% in each case and the average error for the test set of 21 proteins was 5.3%. Importantly, this method allows for an unambiguous determination of the multimeric state of proteins with known molecular weights.
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Citations
Cited by
Accurate assessment of mass, models and resolution by small-angle scattering
Rambo, R. P., & Tainer, J. A. (2013) · Nature · doi:10.1038/nature12070 · Cited by 750
Structural characterization of proteins and complexes using small-angle X-ray solution scattering
Mertens, H. D. T., & Svergun, D. I. (2010) · Journal of Structural Biology · doi:10.1016/j.jsb.2010.06.012 · Cited by 567
Rambo, R. P., & Tainer, J. A. (2011) · Biopolymers · doi:10.1002/bip.21638 · Cited by 533
Small‐angle scattering for structural biology—Expanding the frontier while avoiding the pitfalls
Jacques, D. A., & Trewhella, J. (2010) · Protein Science · doi:10.1002/pro.351 · Cited by 435
Structural and mechanistic insights into regulation of the retromer coat by TBC1d5
Jia, D., Zhang, J.-S., Li, F., Wang, J., Deng, Z., White, M. A., Osborne, D. G., Phillips-Krawczak, C., Gomez, T. S., Li, H., et al. (2016) · Nature Communications · doi:10.1038/ncomms13305 · Cited by 118
Metadata
- Authors
- Fischer, H., de Oliveira Neto, M., Napolitano, H. B., Polikarpov, I., & Craievich, A. F.
- Venue
- Journal of Applied Crystallography
- Published
- 2009-12-01
- Volume
- 43
- Issue
- 1
- Pages
- 101-109
- ISSN
- 0021-8898 (ISSN-L)
- OpenAlex
- W2072435127
- Cited by
- 435
- References
- 0
- FWCI
- 7.82
- Type
- journal article
- Language
- en
- Open access
- closed
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- Fischer, H., de Oliveira Neto, M., Napolitano, H. B., Polikarpov, I., & Craievich, A. F. (2010). Determination of the molecular weight of proteins in solution from a single small-angle X-ray scattering measurement on a relative scale. Journal of Applied Crystallography. https://doi.org/10.1107/s0021889809043076
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
