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Watching a Protein as it Functions with 150-ps Time-Resolved X-ray Crystallography
Abstract
We report picosecond time-resolved x-ray diffraction from the myoglobin (Mb) mutant in which Leu 29 is replaced by Phe (L29Fmutant). The frame-by-frame structural evolution, resolved to 1.8 angstroms, allows one to literally “watch” the protein as it executes its function. Time-resolved mid-infrared spectroscopy of flash-photolyzed L29F MbCO revealed a short-lived CO intermediate whose 140-ps lifetime is shorter than that found in wild-type protein by a factor of 1000. The electron density maps of the protein unveil transient conformational changes far more dramatic than the structural differences between the carboxy and deoxy states and depict the correlated side-chain motion responsible for rapidly sweeping CO away from its primary docking site.
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Citations
Cited by
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Boehr, D. D., Nussinov, R., & Wright, P. E. (2009) · Nature Chemical Biology · doi:10.1038/nchembio.232 · Cited by 2,008
Fenimore, P. W., Frauenfelder, H., McMahon, B. H., & Young, R. D. (2004) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0405573101 · Cited by 524
Ultrafast Spectroscopy: State of the Art and Open Challenges
Maiuri, M., Garavelli, M., & Cerullo, G. (2020) · Journal of the American Chemical Society · doi:10.1021/jacs.9b10533 · Cited by 460
The hit-and-return system enables efficient time-resolved serial synchrotron crystallography
Schulz, E. C., Mehrabi, P., Müller-Werkmeister, H. M., Tellkamp, F., Jha, A., Stuart, W., Persch, E., De Gasparo, R., Diederich, F., Pai, E. F., et al. (2018) · Nature Methods · doi:10.1038/s41592-018-0180-2 · Cited by 111
AlphaFold, allosteric, and orthosteric drug discovery: Ways forward
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Metadata
- Authors
- Schotte, F., Lim, M., Jackson, T. A., Smirnov, A. V., Soman, J., Olson, J. S., Phillips, G. N., Wulff, M., & Anfinrud, P. A.
- Venue
- Science
- Published
- 2003-06-20
- Volume
- 300
- Issue
- 5627
- Pages
- 1944-1947
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W1500927176
- Cited by
- 752
- References
- 0
- FWCI
- 18.54
- Type
- journal article
- Language
- en
- Open access
- closed
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- Schotte, F., Lim, M., Jackson, T. A., Smirnov, A. V., Soman, J., Olson, J. S., Phillips, G. N., Wulff, M., & Anfinrud, P. A. (2003). Watching a Protein as it Functions with 150-ps Time-Resolved X-ray Crystallography. Science. https://doi.org/10.1126/science.1078797
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