Imaging Fluorescence Resonance Energy Transfer as Probe of Membrane Organization and Molecular Associations of GPI-Anchored Proteins
The spatial organization of (GPI)-anchored proteins in cell membranes is a matter of considerable interest. These proteins are thought to be organized into membrane microdomains enriched in GPI-anchored proteins, glycosphingolipids, cholesterol, and some other lipid-modified proteins. Such microdomains have been implicated in membrane trafficking and cell signaling events (reviewed in ref. 1). However, most evidence for the existence of microdomains comes from biochemical studies of isolated membrane fractions (1,2). Microscopy of intact cells has not detected microdomains enriched in GPI-anchored proteins (3, 4, 5); however, these experiments either sample a limited part of the cell surface at high electron-microscope resolution, or an entire cell at low light-microscope resolution.
KeywordsFluorescence Resonance Energy Transfer Energy Transfer Efficiency Lateral Organization Digital Imaging System Donor Fluorescence
- 15.Bastiaens, P. I. H., Wouters, F. S., and Jovin, T. M. (1995) Imaging the molecular state of proteins in cells by fluorescence resonance energy transfer (FRET). Sequential photobleaching of Förster donor-acceptor pairs, in 2nd Hamamatsu International Symposium on Biomolecular Mechanisms and Photonics: Cell-Cell Communications.Google Scholar
- 16.Jovin, T. M. and Arndt-Jovin, D. J. (1989) FRET microscopy: digital imaging of fluorescence resonance energy transfer. Application in cell biology, in Cell Structure and Function by Microspectrofluorimetry, (Kohen, E., Ploem, J. S., and Hirschberg, J. G., eds.), Academic, Orlando, FL, pp. 99–117.Google Scholar