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991.
The G-protein-coupled receptor (GPCR) superfamily represents one of the largest classes of molecules involved in signal transduction
across the plasma membrane. Fluorescence-based approaches have provided valuable insights into GPCR functions such as receptor–receptor
and receptor–ligand interactions, real-time assessment of signal transduction, receptor dynamics on the plasma membrane, and
intracellular trafficking of receptors. This has largely been possible with the use of fluorescent probes such as the green
fluorescent protein (GFP) from the jellyfish Aequoria victoria and its variants. We discuss the potential of fluorescence-based approaches in providing novel information on the membrane
organization and dynamics of the G-protein-coupled serotonin1A receptor tagged to the enhanced yellow fluorescent protein (EYFP).
These authors contributed equally to the work. 相似文献
992.
Lateral diffusion measurements, most commonly accomplished through Fluorescence Photobleaching Recovery (FPR or FRAP), provide
important information on cell membrane molecules' size, environment and participation in intermolecular interactions. However,
serious difficulties arise when these techniques are applied to weakly expressed proteins of either of two types: fusions
of membrane receptors with visible fluorescent proteins or membrane molecules on autofluorescent cells. To achieve adequate
sensitivity in these cases, techniques such as interference fringe FPR are needed. However, in such measurements, cytoplasmic
species contribute to the fluorescence recovery signal and thus yield diffusion parameters not properly representing the small
number of surface molecules. A new method helps eliminate these difficulties. High Probe Intensity (HPI)-FPR measurements
retain the intrinsic confocality of spot measurements to eliminate interference from fluorescent cytoplasmic species. However,
HPI-FPR methods lift the previous requirement that FPR procedures be performed at probe beam intensities low enough to not
induce bleaching in samples during measurements. The high probe intensities now employed provide much larger fluorescence
signals and thus more information on molecular diffusion from each measurement. We report successful measurement of membrane
dynamics by this technique. 相似文献
993.
Fluorimetric Determination of Histamine in Fish Using Micellar Media and Fluorescamine as Labelling Reagent 总被引:1,自引:0,他引:1
Adamou R Coly A Douabalé SE Saleck ML Gaye-Seye MD Tine A 《Journal of fluorescence》2005,15(5):679-688
An analytical method based on the use of fluorescamine to produce a fluorescent derivative with histamine and combined with
micellar-enhanced fluorescence detection of the formed complex is developed for the sensitive and rapid determination of histamine
in fishes. The fluorescence properties of the obtained complex in water and micellar solutions of sodium dodecyl sulfate (SDS),
cetyltrimethylammonium chloride (CTAC) and brij-700 are reported. Physicochemical variables influencing the sensitivity of
the method (pH, micellar, fluorescamine and NaCl relative concentrations) have been optimized. The stability of the formed
complex, as shown by kinetic study, depends on the pH of the solution. Linear calibration curves allowing an effective histamine
determination were established with large linear dynamic range (LDR), and low limits of detection (LOD) between 0.5 and 33
ng mL−1, according to the solvent. Application to the analysis of fish samples (sardines) yielded satisfactory results. The method
seems to be suitable for environmental fish quality control.
Presented in part, at the 39th IUPAC Congress and the 86th Conference of the Canadian Society for Chemistry, August, 10–15,
2003, Ottawa (Canada). 相似文献
994.
In the context, some lanthanide (Eu3+, Tb3+ and Sm3+) complexes with conjugated carboxylic acids (pyridine-carboxylic acids derivatives) have been synthesized and characterized. The low temperature fluorescent spectra for these complexes have been measured at nitrogen atmosphere (77 K), indicating that the central Ln3+ ions locate in an equivalent coordination environment with low symmetry for most of these lanthanide complexes belonging to dimeric or polymeric structure. Therefore, the electronic dipole transition (supersensitive transition) (5D0 → 7F2 for Eu3+, 5D4 → 7F6 for Tb3+, 4G5/2 → 6H9/2 for Sm3+) and magnetic dipole transition (5D0 → 7F1 for Eu3+, 5D4 → 7F5 for Tb3+, 4G5/2 → 6H5/2 for Sm3+) show the regular change in the corresponding split number of fluorescent spectra, which can be realized to predict the fine structure of lanthanide complexes. 相似文献
995.
The emission spectra of naphthalene (NP)–triethylamine (TEA) systems were measured under steady-state illumination conditions in some protic and aprotic solvent-tetrahydrofuran (THF) mixtures. The fluorescence spectrum of the NP–TEA system in THF could be separated into two component bands (band A at 329 nm (fluorescence of NP) and band B at 468 nm (emission from an intermolecular exciplex)). The intensities of bands A and B decreased with increasing solvent polarity. The intensity of band B also decreased owing to the hydrogen-bonding interaction between TEA and protic solvents, but in this case the intensity of band A increased. The decrease in the intensity of band A with increasing solvent polarity is considered to be caused by the enhanced formation of an ion-pair parallel to the formation of an exciplex with increasing solvent polarity. The decrease in the intensity of band B is considered to be caused by the enhanced formation of ion-pair both parallel to and through the formation of the exciplex. The increase in the intensity of band A and the decrease in that of band B upon the addition of protic solvents is caused by the decrease in the concentration of free TEA. Acetonitrile only has a polar effect and trichloroacetic acid only has a hydrogen-bonding (protonation) effect, while alcohols have both the effects. 相似文献
996.
Ortho phthalic anhydride was modified with long chain alcohol (1-docosanol) to its corresponding monodocosyl phthalate (22-Phth). Subsequently, three novel lanthanide (Eu3+, Tb3+, and Dy3+) complexes with the long chain monodocosyl phthalate were synthesized and characterized by elemental analysis and Infrared spectra. The photophysical properties of these complexes were studied in detail with ultraviolet-visible absorption spectra, low temperature phosphorescence spectra and fluorescent spectra. The triplet state energy of 22-Phth was determined to be around 25,000 cm−1 from the maximum phosphorescent peak at 400 nm, suggesting 22-Phth is suitable for the sensitization of the luminescence of Eu3+, Tb3+, and Dy3+. The fluorescence excitation and emission spectra for these lanthanide complexes of the three ligands take agreement with the above predict from energy match principle. 相似文献
997.
We have explored the opportunities for enhanced ratiometric pH sensing using the well-known carboxy seminaphthofluorescein (SNAFL-2) and silver island films (SiFs). Our results show that the metallic surfaces can provide up to a 40-fold increase in probe fluorescence intensity as compared to nonmetallic surfaces with the same probe coverage. However, while the S/N is significantly better for pH sensing, the emission wavelength ratiometric values are similar to that obtained in solution, due to the fact that the emission of both the acidic and basic forms of the probe are enhanced to similar extents. To the best of our knowledge this is the first report of enhanced ratiometric fluorescence sensing on metallic surfaces. 相似文献
998.
999.
Effects of Refractive Index and Viscosity on Fluorescence and Anisotropy Decays of Enhanced Cyan and Yellow Fluorescent Proteins 总被引:1,自引:1,他引:0
The fluorescence lifetime strongly depends on the immediate environment of the fluorophore. Time-resolved fluorescence measurements of the enhanced forms of ECFP and EYFP in water–glycerol mixtures were performed to quantify the effects of the refractive index and viscosity on the fluorescence lifetimes of these proteins. The experimental data show for ECFP and EYFP two fluorescence lifetime components: one short lifetime of about 1 ns and a longer lifetime of about 3.7 ns of ECFP and for EYFP 3.4. The fluorescence of ECFP is very heterogeneous, which can be explained by the presence of two populations: a conformation (67% present) where the fluorophore is less quenched than in the other conformation (33% present). The fluorescence decay of EYFP is much more homogeneous and the amplitude of the short fluorescence lifetime is about 5%. The fluorescence anisotropy decays show that the rotational correlation time of both proteins scales with increasing viscosity of the solvent similarly as shown earlier for GFP. The rotational correlation times are identical for ECFP and EYFP, which can be expected since both proteins have the same shape and size. The only difference observed is the slightly lower initial anisotropy for ECFP as compared to the one of EYFP. 相似文献
1000.