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Organelle-Specific Zinc Detection Using Zinpyr-Labeled Fusion Proteins in Live Cells
Open accessAbstract
A protein labeling approach is employed for the localization of a zinc-responsive fluorescent probe in the mitochondria and in the Golgi apparatus of living cells. ZP1, a zinc sensor of the Zinpyr family, was functionalized with a benzylguanine moiety and thus converted into a substrate (ZP1BG) for the human DNA repair enzyme alkylguaninetransferase (AGT or SNAP-Tag). The labeling reaction of purified glutathione S-transferase tagged AGT with ZP1BG and the zinc response of the resulting protein-bound sensor were confirmed in vitro. The new detection system, which combines a protein labeling methodology with a zinc fluorescent sensor, was tested in live HeLa cells expressing AGT in specific locations. The enzyme was genetically fused to site-directing proteins that anchor the probe onto targeted organelles. Localization of the zinc sensors in the Golgi apparatus and in the mitochondria was demonstrated by fluorescence microscopy. The protein-bound fluorescence detection system is zinc-responsive in living cells.
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Shaner, N. C., Campbell, R. E., Steinbach, P. A., Giepmans, B. N. G., Palmer, A. E., & Tsien, R. Y. (2004) · Nature Biotechnology · doi:10.1038/nbt1037 · Cited by 4,619
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Zhu, H., Fan, J., Du, J., & Peng, X. (2016) · Accounts of Chemical Research · doi:10.1021/acs.accounts.6b00292 · Cited by 984
Advances in fluorescence labeling strategies for dynamic cellular imaging
Dean, K. M., & Palmer, A. E. (2014) · Nature Chemical Biology · doi:10.1038/nchembio.1556 · Cited by 512
Srikun, D., Albers, A. E., Nam, C. I., Iavarone, A. T., & Chang, C. J. (2010) · Journal of the American Chemical Society · doi:10.1021/ja100117u · Cited by 310
Synthesis and application of coumarin fluorescence probes
Sun, X.-Y., Liu, T., Sun, J., & Wang, X.-J. (2020) · RSC Advances · doi:10.1039/c9ra10290f · Cited by 308
Metadata
- Authors
- Tomat, E., Nolan, E. M., Jaworski, J., & Lippard, S. J.
- Venue
- Journal of the American Chemical Society
- Publisher
- American Chemical Society (ACS)
- Published
- 2008-10-31
- Volume
- 130
- Issue
- 47
- Pages
- 15776-15777
- ISSN
- 0002-7863 (ISSN-L)
- 1520-5126
- OpenAlex
- W2025198070
- Cited by
- 202
- References
- 1
- FWCI
- 5.71
- Type
- journal article
- Language
- en
- Open access
- green
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