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Imaging Protein Kinase Cα Activation in Cells
Abstract
Spatially resolved fluorescence resonance energy transfer (FRET) measured by fluorescence lifetime imaging microscopy (FLIM), provides a method for tracing the catalytic activity of fluorescently tagged proteins inside live cell cultures and enables determination of the functional state of proteins in fixed cells and tissues. Here, a dynamic marker of protein kinase Cα (PKCα) activation is identified and exploited. Activation of PKCα is detected through the binding of fluorescently tagged phosphorylation site–specific antibodies; the consequent FRET is measured through the donor fluorophore on PKCα by FLIM. This approach enabled the imaging of PKCα activation in live and fixed cultured cells and was also applied to pathological samples.
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Citations
Cited by
Hunter, T. (2000) · Cell · doi:10.1016/s0092-8674(00)81688-8 · Cited by 2,586
Structural Basis of Protein Kinase C Isoform Function
Steinberg, S. F. (2008) · Physiological Reviews · doi:10.1152/physrev.00034.2007 · Cited by 872
Imaging Protein-Protein Interactions Using Fluorescence Resonance Energy Transfer Microscopy
Kenworthy, A. K. (2001) · Methods · doi:10.1006/meth.2001.1189 · Cited by 532
Imaging biochemistry inside cells
Wouters, F. S., Verveer, P. J., & Bastiaens, P. I. H. (2001) · Trends in Cell Biology · doi:10.1016/s0962-8924(01)01982-1 · Cited by 395
Self-citationThe SUMO modification pathway is involved in the BRCA1 response to genotoxic stress
Morris, J. R., Boutell, C., Keppler, M., Densham, R., Weekes, D., Alamshah, A., Butler, L., Galanty, Y., Pangon, L., Kiuchi, T., et al. (2009) · Nature · doi:10.1038/nature08593 · Cited by 375
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Metadata
- Authors
- Ng, T., Squire, A., Hansra, G., Bornancin, F., Prevostel, C., Hanby, A., Harris, W., Barnes, D., Schmidt, S., Mellor, H., Bastiaens, P. I. H., & Parker, P. J.
- Venue
- Science
- Published
- 1999-03-26
- Volume
- 283
- Issue
- 5410
- Pages
- 2085-2089
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W2125551363
- Cited by
- 287
- References
- 0
- FWCI
- 18.63
- Type
- journal article
- Language
- en
- Open access
- closed
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- Ng, T., Squire, A., Hansra, G., Bornancin, F., Prevostel, C., Hanby, A., Harris, W., Barnes, D., Schmidt, S., Mellor, H., Bastiaens, P. I. H., & Parker, P. J. (1999). Imaging Protein Kinase Cα Activation in Cells. Science. https://doi.org/10.1126/science.283.5410.2085
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