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Title: | New fluorescent octadecapentaenoic acids as probes of lipid membranes and protein-lipid interactions |
Authors: | Mateo, C. Reyes; Souto, André A.; Amat-Guerri, Francisco CSIC; Acuña, A. Ulises CSIC | Keywords: | Lipid membranes Protein-lipid interactions Fluorescent acids Octadecapentaenoic acid |
Issue Date: | 1-Oct-1996 | Publisher: | Rockefeller University Press | Citation: | Biophysical Journal 71, 1428-1439 (1996) | Abstract: | The chemical and spectroscopic properties of the new fluorescent acids all(E)-8, 10, 12, 14, 16-octadecapentaenoic acid (t-COPA) and its (8Z)-isomer (c-COPA) have been characterized in solvents of different polarity, synthetic lipid bilayers, and lipid/protein systems. These compounds are reasonably photostable in solution, present an intense UV absorption band (epsilon(350 nm) approximately 10(5) M(-1) cm(-1)) strongly overlapped by tryptophan fluorescence and their emission, centered at 470 nm, is strongly polarized (r(O) = 0.385 +/- 0.005) and decays with a major component (85%) of lifetime 23 ns and a faster minor one of lifetime 2 ns (D,L-alpha-dimyristoylphosphatidylcholine (DMPC), 15 degrees C). Both COPA isomers incorporate readily into vesicles and membranes (K(p) approximately 10(6)) and align parallel to the lipids. t-COPA distributes homogeneously between gel and fluid lipid domains and the changes in polarization accurately reflect the lipid T(m) values. From the decay of the fluorescence anisotropy in spherical bilayers of DMPC and POPC it is shown that t-COPA also correctly reflects the lipid order parameters, determined by 2H NMR techniques. Resonance energy transfer from tryptophan to the bound pentaenoic acid in serum albumin in solution, and from the tryptophan residues of gramicidin in lipid bilayers also containing the pentaenoic acid, show that this probe is a useful acceptor of protein tryptophan excitation, with R(O) values of 3034 A. | Publisher version (URL): | http://dx.doi.org/10.1016/S0006-3495(96)79419-5 | URI: | http://hdl.handle.net/10261/11913 | DOI: | 10.1016/S0006-3495(96)79419-5 | ISSN: | 0006-3495 |
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