Fluorescent Biomembrane Probe for Ratiometric Detection of Apoptosis Export

@article{citeulike:1128589, abstract = {Abstract: Herein, we developed the first ratiometric fluorescent probe for apoptosis detection. This probe incorporates selectively into the outer leaflet of the cell plasma membrane and senses the loss of the plasma membrane asymmetry occurring during the early steps of apoptosis. The high specificity to the plasma membranes was achieved by introduction into the probe of a membrane anchor, composed of a zwitterionic group and a long (dodecyl) hydrophobic tail. The fluorescence reporter of this probe is 4'-(diethylamino)-3-hydroxyflavone, which exhibits excited-state intramolecular proton transfer (ESIPT), resulting in two-band emission highly sensitive to the lipid composition of the biomembranes. Fluorescence spectroscopy, flow cytometry, and microscopy measurements show that the ratio of the two emission bands of the probe changes dramatically in response to apoptosis. This response reflects the changes in the lipid composition of the outer leaflet of the cell plasma membrane because of the exposure of the anionic phospholipids from the inner leaflet at the early steps of apoptosis. Being ratiometric, the response of the new probe can be easily quantified on an absolute scale. This allows monitoring by laser scanning confocal microscopy the degree and spatial distribution of the apoptotic changes at the cell plasma membranes, a feature that can be hardly achieved with the commonly used fluorescently labeled annexin V assay.}, address = {Contribution from the Photophysique des Interactions Mol\'{e}culaires, UMR 7175 CNRS, Institut Gilbert Laustriat, Facult\'{e} de Pharmacie, Universit\'{e} Louis Pasteur, Strasbourg 1, 74, Route du Rhin, 67401 Illkirch Cedex, France, Department of Physics, Kyiv National Taras Shevchenko University, 01033 Kyiv, Ukraine, Unit\'{e} 143 INSERM et Institute d'Hematologie et d'Immunologie Universit\'{e} Louis Pasteur, Strasbourg 67085, France, and Palladin Institute of Biochemistry, 01030 Kyiv, Ukraine}, author = {Shynkar, V. V. and Klymchenko, A. S. and Kunzelmann, C. and Duportail, G. and Muller, C. D. and Demchenko, A. P. and Freyssinet, J. M. and Mely, Y.}, citeulike-article-id = {1128589}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/ja068008h}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/ja068008h}, day = {21}, doi = {10.1021/ja068008h}, journal = {J. Am. Chem. Soc.}, keywords = {biophysics, dual-emission, esipt, readout}, month = {February}, number = {7}, pages = {2187--2193}, posted-at = {2007-02-28 00:38:45}, priority = {1}, title = {Fluorescent Biomembrane Probe for Ratiometric Detection of Apoptosis}, url = {http://dx.doi.org/10.1021/ja068008h}, volume = {129}, year = {2007} }

TY - JOUR ID - citeulike:1128589 L3 - citeulike-article-id:1128589 N2 - Abstract: Herein, we developed the first ratiometric fluorescent probe for apoptosis detection. This probe incorporates selectively into the outer leaflet of the cell plasma membrane and senses the loss of the plasma membrane asymmetry occurring during the early steps of apoptosis. The high specificity to the plasma membranes was achieved by introduction into the probe of a membrane anchor, composed of a zwitterionic group and a long (dodecyl) hydrophobic tail. The fluorescence reporter of this probe is 4'-(diethylamino)-3-hydroxyflavone, which exhibits excited-state intramolecular proton transfer (ESIPT), resulting in two-band emission highly sensitive to the lipid composition of the biomembranes. Fluorescence spectroscopy, flow cytometry, and microscopy measurements show that the ratio of the two emission bands of the probe changes dramatically in response to apoptosis. This response reflects the changes in the lipid composition of the outer leaflet of the cell plasma membrane because of the exposure of the anionic phospholipids from the inner leaflet at the early steps of apoptosis. Being ratiometric, the response of the new probe can be easily quantified on an absolute scale. This allows monitoring by laser scanning confocal microscopy the degree and spatial distribution of the apoptotic changes at the cell plasma membranes, a feature that can be hardly achieved with the commonly used fluorescently labeled annexin V assay. IS - 7 JF - J. Am. Chem. Soc. AD - Contribution from the Photophysique des Interactions Moléculaires, UMR 7175 CNRS, Institut Gilbert Laustriat, Faculté de Pharmacie, Université Louis Pasteur, Strasbourg 1, 74, Route du Rhin, 67401 Illkirch Cedex, France, Department of Physics, Kyiv National Taras Shevchenko University, 01033 Kyiv, Ukraine, Unité 143 INSERM et Institute d'Hematologie et d'Immunologie Université Louis Pasteur, Strasbourg 67085, France, and Palladin Institute of Biochemistry, 01030 Kyiv, Ukraine EP - 2193 TI - Fluorescent Biomembrane Probe for Ratiometric Detection of Apoptosis VL - 129 SP - 2187 KW - biophysics KW - dual-emission KW - esipt KW - readout AU - Shynkar, VV AU - Klymchenko, AS AU - Kunzelmann, C AU - Duportail, G AU - Muller, CD AU - Demchenko, AP AU - Freyssinet, JM AU - Mely, Y PY - 2007/02/21/ UR - http://dx.doi.org/10.1021/ja068008h ER -