Binding interaction of differently charged fluorescent probes with egg yolk phosphatidylcholine and the effect of ß-cyclodextrin on the lipid-probe complexes: a fluorometric investigation.

Interaction of cationic phenosafranin (PSF), anionic 8-anilino-1-naphthalene sulfonate (ANS) and non-ionic nile red (NR) have been studied with the zwitterionic phospholipid, egg yolk L-α-phosphatidylcholine (EYPC). The study reveals discernible binding interactions of the three fluorescent probes with the EYPC lipid vesicle. Once the binding of the probes with the lipid is established, the effect of cyclic oligosaccharide, ß-cyclodextrin (ß-CD), on these lipid bound probes has been investigated. Different fluorometric techniques suggest that addition of ß-CD to the probe-lipid complexes leads to the release of the probes from the lipid medium through the formation of probe-ß-CD inclusion complexes. A competitive binding of the probes between ß-cyclodextrin and the lipid is ascribed to be responsible for the effect. This provides an easy avenue for the removal of the probe molecules from the lipid environment. Extension of this work with drug molecules in cell membranes is expected to give rise to a strategy for the removal of adsorbed drugs from the cell membranes by the use of non-toxic ß-cyclodextrin.

Competitive binding of nile red between lipids and ß-cyclodextrin.

Fluorometric measurements are exploited to explore the binding interactions of nile red (NR) with anionic lipid dimyristoyl-L-α-phosphatidylglycerol (DMPG), zwitterionic lipid dimyristoyl-L-α-phosphatidylcholine (DMPC) as well as neutral cyclic oligosaccharide ß-cyclodextrin (ß-CD) solutions. The binding constants are found to be quite high and comparable (within a factor of five). Series of spectral techniques like steady state fluorescence and fluorescence anisotropy study, micropolarity study, quenching study and time resolved experiments reveal that the addition of ß-CD to the probe-lipid complexes results in weakening of the lipid-probe interaction and formation of probe-ß-CD inclusion complexes leading to the removal of some of the probe (NR) molecules from the lipid environments. The extent of removal of NR is, however, more from DMPG than DMPC lipid. The phenomena are explained from the concept of competitive binding of the probe between the lipids and ß-CD. Since lipids are the principal constituents of the cell walls, the work might make a foundation for the possible removal of excess of molecules like nile red adsorbed on the cell walls.

Multiple emissions of α-naphthil: fluorescence from S2 state.

Multiple emissions of α-naphthil are reported in solution phase at room temperature as well as in ethanol glassy matrix at 77 K, frozen under different conditions of illumination. Steady state and time-resolved fluorometric measurements are undertaken to assign the observed emissions to the different species in terms of geometry and electronic states of the fluorophore. The unexplored emission band peaking at ∼423 nm has been ascribed to originate from the S2 electronic state. Exhaustive experiments reveal that while the trans conformer of α-naphthil gives only the S1 emission, the conformation with a geometry close to the ground state one emits from both S1 and S2 electronic states depending on the excitation wavelength.

Spectroscopic exploration of mode of binding of ctDNA with 3-hydroxyflavone: a contrast to the mode of binding with flavonoids having additional hydroxyl groups.

Binding interaction of 3-hydroxyflavone (3HF), a bioactive flavonoid, with calf-thymus DNA (ctDNA) has been explored exploiting various experimental techniques. The dual fluorescence of 3HF resulting from the excited state intramolecular proton transfer (ESIPT) is modified remarkably upon binding with the biomacromolecule. The determined binding constant, fluorescence quenching experiment, circular dichroism (CD) study, comparative binding study with the known intercalative binder ethidium bromide and thermometric experiment relating to the helix melting of ctDNA confirm the groove binding of 3HF with the DNA. This is in contrast to two other members of the flavonoid group, namely, fisetin and quercetin, where the bindings are established to be intercalative. The structural difference of 3HF from the other two probes with respect to the absence/presence of the additional hydroxyl groups is ascribed to be responsible for the difference in the mode of binding.

Multiple emissions of benzil at room temperature and 77 K and their assignments from ab initio quantum chemical calculations.

Multiple emissions have been observed from benzil under different conditions in solutions at room temperature as well as in low temperature glass matrices at 77 K. Low temperature emission has been monitored in rigid matrices frozen under different conditions of illumination. Steady state and time-resolved results together with the ab initio quantum chemical calculations provide, for the first time, the assignments of the different fluorescence bands to the different geometries and∕or electronic states of the fluorophore molecule. It is revealed that the skew form of benzil emits from the first (S(1)) as well as the second excited singlet (S(2)) states depending on the excitation wavelength, while the relaxed transplanar conformer fluoresces only from the S(1) state. The yet unexplored emission band peaking at around 360 nm has been assigned to originate from the S(2) state. Ab initio calculations using the density functional theory at B3LYP∕6-31G∗∗ level corroborate well with the experimental observations.

Studies of Triton X-165-beta-cyclodextrin interactions using both extrinsic and intrinsic fluorescence.

The interaction of beta-cyclodextrin with the non-ionic micelle-forming surfactant Triton X-165 (TX-165) has been studied using steady state fluorescence and fluorescence anisotropy techniques. Both extrinsic and intrinsic fluorescence have been exploited for the purpose. Phenosafranin (PSF), a cationic phenazinium dye, has been used as the extrinsic probe while fluorescence of TX-165 has served as the intrinsic one. PSF shows discernible interactions with both TX-165 and beta-CD. The experimental results reveal that the extent of interaction of PSF with TX-165 is greater than with beta-CD. However, addition of beta-CD to a micellar solution of TX-165 containing PSF leads to a disruption of the micelles whereby the fluorophore is released from the micellar environment to the bulk aqueous phase. It has been substantiated that an inclusion complex is formed between the non-ionic surfactant and the cyclodextrin. A 1:1 stoichiometry of the TX-165-beta-CD inclusion complex has been proposed. Such a complexation between TX-165 and beta-CD results in an inhibition in the micellization process of TX-165 leading to an enhancement in the apparent CMC value. The inferences are drawn from a series of experiments, viz., binding studies, determination of micropolarity, heavy-ion quenching studies and steady state fluorescence anisotropy experiments monitoring both extrinsic and intrinsic fluorescences.