We have systematically investigated the effect of various alkali metal ions with negatively charged phospholipid membranes. Size distributions of large unilamellar vesicles have been confirmed using dynamic light scattering. Zeta potential and effective charges per vesicle in the presence of various alkali metal ions have been estimated from the measured electrophoretic mobility. We have determined the intrinsic binding constant from the zeta potential using electrostatic double layer theory. The reasonable and consistent value of the intrinsic binding constant of Na(+), found at moderate NaCl concentration (10-100 mM), indicates that the Gouy-Chapman theory cannot be applied for very high (> 100mM) and very low (< 10 mM) electrolyte concentrations. The isothermal titration calorimetry study has revealed that the net binding heat of interaction of the negatively charged vesicles with monovalent alkali metal ions is small and comparable to those obtained from neutral phosphatidylcholine vesicles. The overall endothermic response of binding heat suggests that interaction is primarily entropy driven. The entropy gain might arise due to the release of water molecules from the hydration layer vicinity of the membranes. Therefore, the partition model which does not include the electrostatic contribution suffices to describe the interaction. The binding constant of Na(+) (2.4 ± 0.1 M(-1)), obtained from the ITC, is in agreement with that estimated from the zeta potential (-2.0 M(-1)) at moderate salt concentrations. Our results suggest that hydration dynamics may play a vital role in the membrane solution interface which strongly affects the ion-membrane interaction.
A comparative
binding interaction of toluidine blue O (TBO) and
methylene blue (MB) with lysozyme was investigated by multifaceted
biophysical approaches as well as from the aspects of in silico biophysics.
The bindings were static, and it occurred via ground-state complex
formation as confirmed from time-resolved fluorescence experiments.
From steady-state fluorescence and anisotropy, binding constants were
calculated, and it was found that TBO binds more effectively than
MB. Synchronous fluorescence spectra revealed that binding of dyes
to lysozyme causes polarity changes around the tryptophan (Trp) moiety,
most likely at Trp 62 and 63. Calorimetric titration also depicts
the higher binding affinity of TBO over MB, and the interactions were
exothermic and entropy-driven. In silico studies revealed the potential
binding pockets in lysozyme and the participation of residues Trp
62 and 63 in ligand binding. Furthermore, calculations of thermodynamic
parameters from the theoretical docking studies were in compliance
with experimental observations. Moreover, an inhibitory effect of
these dyes to lysozyme fibrillogenesis was examined, and the morphology
of the formed fibril was scanned by atomic force microscopy imaging.
TBO was observed to exhibit higher potential in inhibiting the fibrillogenesis
than MB, and this phenomenon stands out as a promising antiamyloid
therapeutic strategy.
We report the generation of simple condensates of short peptides with ATP, which are spatiotemporally formed under dissipative conditions and temporally modulate a secondary redox reaction catalyzed by the entrapped protein.
We have systematically investigated the effect of counterions on the interaction of the Na+ ion with phospholipid membranes using dynamic light scattering, zeta potential, isothermal titration calorimetry and fluorescence spectroscopy techniques.
Cationic lipids have attracted much attention because of their potential for biomedical applications, such as gene delivery. The gene transfection efficiency of cationic lipids is greatly influenced by the counterions as well as salt ions. We have systematically investigated the interaction of different monovalent sodium salts with positively charged membrane, composed of 1,2-dioleoyl- sn-glycero-3-phosphocholine (DOPC)/1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and DOTAP, using dynamic light scattering, zeta potential, isothermal titration calorimetry (ITC), and fluorescence spectroscopy techniques. Our results reveal that the affinity of anions with cationic membranes follows the sequence I ≫ Br > Cl according to descending order of their sizes and is consistent with the Hofmeister series. Interestingly, the electrostatic behavior of the DOTAP membrane in the presence of monovalent anions differs significantly from the DOPC/DOTAP membrane. This difference is due to the strong interplay between phosphocholine and trimethylammonium-propane (TAP) headgroups leading to the reorientation of the TAP group in the membrane. The binding constant of anions, derived from zeta potential and ITC is in agreement with the affinity of anions mentioned above. Among all anions, I shows strongest affinity, as evidenced from the rapid increase in hydrodynamic radius which eventually leads to the formation of large aggregates. The fluorescence spectroscopy of a lypophilic probe Nile red in the presence of cationic vesicles containing ions complements the I adsorption onto the membrane. Nonlinear Stern-Volmer plot, consisting of accessible and inaccessible Nile red to I is consistent with the zeta potential as well as ITC results.
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