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Chemical Probes for the Rapid Detection of Fatty-Acylated Proteins in Mammalian Cells
Abstract
The attachment of lipids onto proteins modulates the activity of proteins in many biological settings. The analysis of protein lipidation, however, is challenging due to the relatively few methods for the detection of lipid-modified proteins. Here we describe the synthesis of ω-azido-fatty acids as non-radioactive chemical probes for the rapid visualization of fatty-acylated proteins in mammalian cells. Following metabolic installation of the ω-azido-fatty acids onto target proteins by cellular enzymes, fatty-acylated proteins are selectively biotinylated with a phosphine−biotin reagent via the Staudinger ligation and visualized by streptavidin blotting. Depending on the chain length of the ω-azido-fatty acids, N-myristoylated and S-palmitoylated proteins can be visualized selectively in cell lysates and on specific proteins. These chemical probes provide new tools to analyze fatty acylation of proteins in living cells.
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Cited by
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Lang, K., & Chin, J. W. (2014) · Chemical Reviews · doi:10.1021/cr400355w · Cited by 1,035
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McKay, C. S., & Finn, M. G. (2014) · Chemistry & Biology · doi:10.1016/j.chembiol.2014.09.002 · Cited by 768
Protein post-translational modifications in bacteria
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Metadata
- Authors
- Hang, H. C., Geutjes, E.-J., Grotenbreg, G., Pollington, A. M., Bijlmakers, M. J., & Ploegh, H. L.
- Venue
- Journal of the American Chemical Society
- Publisher
- American Chemical Society (ACS)
- Published
- 2007-02-17
- Volume
- 129
- Issue
- 10
- Pages
- 2744-2745
- ISSN
- 0002-7863 (ISSN-L)
- 1520-5126
- OpenAlex
- W1980541667
- Cited by
- 205
- References
- 0
- FWCI
- 10.55
- Type
- journal article
- Language
- en
- Open access
- closed
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- Hang, H. C., Geutjes, E.-J., Grotenbreg, G., Pollington, A. M., Bijlmakers, M. J., & Ploegh, H. L. (2007). Chemical Probes for the Rapid Detection of Fatty-Acylated Proteins in Mammalian Cells. Journal of the American Chemical Society. https://doi.org/10.1021/ja0685001
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