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Labeling and quantifying sites of protein palmitoylation
Open accessAbstract
As a reversible posttranslational modification, protein palmitoylation has the potential to regulate the trafficking and function of a variety of proteins. However, the extent, function, and dynamic nature of palmitoylation are poorly resolved because of limitations in assay methods. Here, we introduce methods where hydroxylamine-mediated cleavage of the palmitoyl-thioester bond generates a free sulfhydryl, which can then be specifically labeled with sulfhydryl-reactive reagents. This methodology is more sensitive and allows for quantitative estimates of palmitoylation. Unlike other techniques used to assay posttranslational modifications, the techniques we have developed can label all sites of modification with a variety of probes, radiolabeled or nonradioactive, and can be used to assay the palmitoylation of proteins expressed in vivo in brain or other tissues.
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Cited by
Global Analysis of Protein Palmitoylation in Yeast
Roth, A. F., Wan, J., Bailey, A. O., Sun, B., Kuchar, J. A., Green, W. N., Phinney, B. S., Yates, J. R., & Davis, N. G. (2006) · Cell · doi:10.1016/j.cell.2006.03.042 · Cited by 543
Self-citationNeural palmitoyl-proteomics reveals dynamic synaptic palmitoylation
Kang, R., Wan, J., Arstikaitis, P., Takahashi, H., Huang, K., Bailey, A. O., Thompson, J. X., Roth, A. F., Drisdel, R. C., Mastro, R., et al. (2008) · Nature · doi:10.1038/nature07605 · Cited by 537
Self-citationPalmitoylation of Ligands, Receptors, and Intracellular Signaling Molecules
Resh, M. D. (2006) · Science's STKE · doi:10.1126/stke.3592006re14 · Cited by 414
Rho Family GTPase modification and dependence on CAAX motif-signaled posttranslational modification
Roberts, P. J., Mitin, N., Keller, P. J., Chenette, E. J., Madigan, J. P., Currin, R. O., Cox, A. D., Wilson, O., Kirschmeier, P., & Der, C. J. (2008) · Journal of Biological Chemistry · doi:10.1074/jbc.m800882200 · Cited by 362
The intracellular dynamic of protein palmitoylation
Salaun, C., Greaves, J., & Chamberlain, L. H. (2010) · Journal of Cell Biology · doi:10.1083/jcb.201008160 · Cited by 355
Metadata
- Authors
- Drisdel, R. C., & Green, W. N.
- Venue
- BioTechniques
- Publisher
- Informa UK Limited
- Published
- 2004-02
- Volume
- 36
- Issue
- 2
- Pages
- 276-285
- ISSN
- 0736-6205 (ISSN-L)
- 1940-9818
- OpenAlex
- W35736013
- Cited by
- 325
- References
- 0
- FWCI
- 3.50
- Type
- journal article
- Language
- en
- Open access
- gold
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