Blue- and Red-Shifting C-H⋯O Hydrogen Bonds of Cyclic Ethers with Haloforms: Effect of Ring-Size and Consistency with Bent's Rule.

A DFT-based computational study is carried out to delve into the interplay between hyperconjugation and rehybridization effects underlying the formation of blue- or red-shifting H-bonds (HBs) in 1 : 1 complexes of cyclic ethers (HB acceptor) of varying ring-size with haloforms, CHF3 and CHCl3 (HB donor). The calculations reveal that with decreasing angular strain (increasing ring-size) of the cyclic ethers, the extent of blue-shift increases for 1 : 1 complexes with CHF3, while a reverse sequence is observed with CHCl3, eventually leading to a red-shifting HB in the oxepane : CHCl3 complex. It is noted that the trend in the shift of C-H stretching fundamental is not mirrored by the C-H bond length or interaction energies for both the systems studied, that is, the low sensitivity of the changes on the strain on the O-atom of HB acceptor (cyclic ethers) is to be emphasized.

Exploring the Nucleobase-Specific Hydrophobic Interaction of Cryptolepine Hydrate with RNA and Its Subsequent Sequestration.

The study of the interactions of drug molecules with genetic materials plays a key role underlying the development of new drugs for many life-threatening diseases in pharmaceutical industries. Understanding their fundamental base-specific and/or groove-binding interaction is crucial to target the genetic material with an external drug, which can pave the way to curing diseases related to the genetic material. Here, we studied the interaction of cryptolepine hydrate (CRYP) with RNA under physiological conditions knowing the antimalarial and anticancer activities of the drug. Our experiments explicitly demonstrate that CRYP interacts with the guanine- and adenine-rich region within the RNA duplex. The pivotal role of the hydrophobic interaction governing the interaction is substantiated by temperature-dependent isothermal titration calorimetry experiments and spectroscopic studies. Circular dichroism study underpins a principally intercalative mode of binding of CRYP with RNA. This interaction is found to be drastically affected in the presence of magnesium salt, which has a strong propensity to coordinate with RNA nucleobases, which can in turn modulate the interaction of the drug with RNA. The temperature-dependent calorimetric results substantiate the occurrence of entropy-enthalpy compensation, which enabled us to rule out the possibility of groove binding of the drug with RNA. Furthermore, our results also show the application of host-guest chemistry in sequestering the RNA-bound drug, which is crucial to the development of safer therapeutic applications.

Contrasting Thermodynamics Governs the Interaction of 3-Hydroxyflavone with the N-Isoform and B-Isoform of Human Serum Albumin.

Herein we report the interaction of 3-hydroxyflavone (3HF) with various isomeric forms of Human Serum Albumin (HSA), namely, the N-isoform (or native HSA at pH 7.4) and the B-isoform (at pH 9.2). Spectroscopic signatures of 3HF reveal that the interaction of 3HF with the N-isoform of HSA results in significant lowering of absorbance of the neutral species (λabs ∼ 345 nm) with concomitant increase of the anionic species (λabs ∼ 416 nm) whereas interaction with the B-isoform of HSA leads to selective enhancement of absorbance of the anionic species. The fluorescence profile of 3HF displays marked increase of intensity of the proton transferred tautomer (λem ∼ 538 nm) as well as the anionic species (λem ∼ 501 nm) for both the forms of the protein. However, analyses of the associated thermodynamics through temperature-dependent isothermal titration calorimetric (ITC) indicate that the interaction of 3HF with the N-isoform of HSA is more enthalpic in the lower temperature limit while the entropy contribution predominates in the higher temperature limit. Consequently, the 3HF-HSA (N-isoform at pH 7.4) interaction reveals an unusual thermodynamic signature of a positive heat capacity change (ΔCp = 3.84 kJ mol-1K-1) suggesting the instrumental role of hydrophobic hydration. On the contrary, the 3HF-HSA (B-isoform at pH 9.2) interaction shows qualitatively reverse effect. Consequently, the interaction is found to be characterized by an enthalpy-dominated hydrophobic effect (negative heat capacity change, ΔCp = -1.15 kJ mol-1K-1) which is rationalized on the basis of the nonclassical hydrophobic effect.

Effect of temperature and salts on niosome-bound anti-cancer drug along with disruptive influence of cyclodextrins.

Encapsulation of a persuasive anticancer drug (Sanguinarine, SGR) within microheterogeneous environment of niosome has been investigated. Utilizing steady-state and time-resolved spectroscopic methods the effects of extrinsically added salts and temperature on the photophysical properties of niosome-bound bio-active drug have been explored thoroughly. The prototropic (alkanolamine⇌ iminium) equilibrium of SGR is found to be preferentially favored toward the neutral form inside the hydrophobic interior of niosome. With addition of salts and increment of temperature the reverse tendency of stabilization of the cationic species is observed which can be explained on the basis of degree of water penetration of water molecules to the hydration layer of niosome. Furthermore, drug sequestration has been investigated via disruption of niosome applying cyclodextrins (CDs). Exploration of the effect of CDs (ß-CD and γ-CD) on the niosome aids to have knowledge of the effect of CDs on cell membrane. In addition, the differential rotational relaxation behavior of SGR at various environmental circumstances has been observed to substantiate with other experimental results.

How is the interaction of a chloride channel blocker with phospholipids influenced by divalent metal ions? Effect of unsaturation on the lipid side chain.

The present study reveals the effect of various divalent ions (Ca2+, Mg2+and Zn2+) on the binding interaction of a prospective chloride channel blocker, 9-methylanthroate (9MA), with liposome membranes, namely, dimyristoylphosphatidylcholine (DMPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). The liposome membranes DMPC and POPC differ in the unsaturation of the side-chain. The drug (9MA) is found to experience a greater degree of partitioning into the POPC lipid bilayer (containing unsaturated side-chain) in comparison to DMPC (containing saturated side-chain). The stronger 9MA-POPC binding interaction is found to be only nominally perturbed by the presence of metal salts. On the contrary, the 9MA-DMPC binding interaction is found to be significantly perturbed by the presence of metal salts and is manifested on the environment-responsive spectroscopic properties of the drug. The steady-state and picosecond-resolved fluorescence spectroscopic results reveal the effect of metal ions on DMPC bilayer to follow the trend Ca2+ < Mg2+ < Zn2+. This is also quantified by evaluating the partition coefficient of the drug into DMPC lipid in the presence of various divalent ions which is found to follow the same sequence. The degree of penetration of these cations has been rationalized on the basis of adsorption of cations on DMPC headgroup region resulting in dehydration of the headgroup along with shrinking of it.

Revealing the dynamics and energetics of interaction of a cationic biological photosensitizer within a bile salt aggregate.

The present investigation reports a detailed characterization of the interaction of a cationic photosensitizer, phenosafranin (PSF) with sodium deoxycholate (NaDC) bile salt aggregates based on spectroscopic and calorimetric techniques. Our explicit spectroscopic results not only establish the occurrence of PSF-NaDC binding interaction, but also reveal marked lowering of micropolarity at the interaction site (ET(30) = 55.97 kcal mol-1 in the presence of NaDC as compared to ET(30) = 63.1 kcal mol-1 in bulk aqueous buffer). A thorough mathematical analysis of the fluorescence depolarization results based on the two-step and wobbling in cone model yields critical insight into the complex rotational relaxation dynamics of the bound drug. The impartation of motional restriction on the PSF molecules within the bile salt aggregates is evidenced from enhancement of average rotational correlation time from <τr> = 136 ps in aqueous buffer to 1.11 ns with added NaDC (8.0 mM). This is further supported from a high value of the generalized order parameter (S = 0.81) as well as the diffusion coefficient (Dw = 1.40 × 1012 s-1). Furthermore, our extensive calorimetric investigation unveils the complicated thermodynamics of the interaction process in terms of predominant entropic contribution over the enthalpic part in the lower temperature regime (TΔS = 18.84 ±â€¯1.13 kJ mol-1, ΔH = -5.82 ±â€¯0.35 kJ mol-1 at 288 K) with subsequent reversal of the relative contributions with increasing temperature (TΔS = 7.54 ±â€¯0.39 kJ mol-1, ΔH = - 17.09 ±â€¯0.90 kJ mol-1 at 318 K). The instrumental role of the hydrophobic effect underlying the PSF-NaDC interaction is characterized by a negative heat capacity change (ΔCp = -364 J mol-1 K-1). An intriguing thermodynamic feature in terms of enthalpy-entropy compensation (with increasing temperature ΔG remains almost constant while ΔH and TΔS vary significantly) aptly corroborates the aforesaid argument and establishes an appreciable hydrophobic contribution to the overall binding energies.

Unsaturation of the phospholipid side-chain influences its interaction with cyclodextrins: A spectroscopic exploration using a phenazinium dye.

The interaction of a cationic photosensitizer Safranin-O with liposome membranes having similar surface charge (negative) but differing in the presence of saturation on the lipid side-chain has been studied. To this end, dimyristoyl-l-R-phosphatidylglycerol (DMPG) and 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DOPG) phospholipids were employed to prepare small unilamellar vesicles. The dye is found to bind in the headgroup region of both the liposome membranes with significantly higher affinity to DOPG lipid containing unsaturated side chain. The effects of various cyclodextrins (CDs) on the stability of the probe-bound liposome membranes have also been investigated using steady-state and picosecond-resolved fluorescence as well as dynamic light scattering techniques. The modulations of the fluorescence properties of the lipid-bound dye were exploited to rationalize the membrane destabilization following interaction with the cyclodextrins. Experimental results reveal the selective interaction of DMPG membrane with CDs leading to rupture of the integrated structure of the liposome units accompanying release of the bound probe to the bulk aqueous phase. On the contrary, no discernible interaction of the CDs was observed with DOPG liposome membrane. Our results also show the differential extents of interaction of various CDs (α-CD, ß-CD, methyl-ß-CD, and γ-CD) with DMPG leading to varying degrees of release of the bound-dye molecule.

Binding of norharmane with RNA reveals two thermodynamically different binding modes with opposing heat capacity changes.

The binding interaction of a prospective anti-cancer photosensitizer, norharmane (NHM, 9H-pyrido[3,4-b]indole) with double stranded RNA reveals a primarily intercalative mode of binding. Steady-state and time-resolved fluorescence spectroscopic results demonstrate the occurrence of drug-RNA binding interaction as manifested through environment-sensitive prototropic equilibrium of NHM. However, the key finding of the present study lies in unraveling the complexities in the NHM-RNA binding thermodynamics. Isothermal Titration Calorimetry (ITC) results reveal the presence of two thermodynamically different binding modes for NHM. An extensive temperature-dependence investigation shows that the formation of Complex I is enthalpically (ΔHI < 0) as well as entropically (TΔSI > 0) favored with the enthalpic (entropic) contribution being increasingly predominant in the higher (lower) temperature regime. On the contrary, the formation of Complex II reveals a predominantly enthalpy-driven signature (ΔHI < 0) along with unfavorable entropy change (TΔSI < 0) with gradually decreasing enthalpic contribution with temperature. Such differential dependences of ΔHI and ΔHII on temperature subsequently lead to opposing heat capacity changes underlying the formation of Complex I and II (ΔCpI<0andΔCpII>0). A negative ΔCp underpins the pivotal role of 'hydrophobic effect' (release of ordered water molecules) for the formation of Complex I, while a positive ΔCp marks the thermodynamic hallmark for 'hydrophobic hydration' (solvation of hydrophobic (or nonpolar) molecular surfaces in aqueous medium) for formation of Complex II. A detailed investigation of the effect of ionic strength enables a component analysis of the total free energy change (ΔG).

Interaction of a cationic amphiphile with monomeric and polymeric electrolytes: From morphological transition to associative phase separation.

The discrete effects of a series of structurally divergent monomeric viz. Sodium Chloride (NaCl), Tetra-butyl Ammonium Chloride (TBAC) and Sodium Benzoate (NaBz) and polymeric viz. Sodium Polystyrene Sulfonate (NaPSS) electrolytes towards the morphological and/or aggregation properties of Octadecyl-trimethyl Ammonium Bromide (OTAB) micelles have been quantified spectroscopically by means of the modulations of the absorption and emission spectral properties of an extrinsic anthracene-based probe 9-methyl anthroate (9-MA) within the concerned media. Further corroboration of the spectroscopic results was acquired from the non-invasive dynamic light scattering technique. The qualitatively similar mode of action of all the monomeric salts has been explained on the basis of the archetypal Israelachvili model whereas the corresponding extent of the morphological transition of the micelles, which is found to follow the order NaBz > NaCl > TBAC, has been explained invoking the co-sphere overlap model. Conversely, to explain the aggregation behaviour of the micelles in the presence of the polymeric electrolyte, a two-step model has been formulated. According to this model, at the low concentration regime, the polymeric salt is found to only neutralize the surface charge of the micelles inducing micellar growth; whereas further increment in the concentration of the polymer assists the hydrophobic association between the micelles leading to the formation of larger aggregates, eventually causing a phase separation.

Binding interaction of phenazinium-based cationic photosensitizers with human hemoglobin: Exploring the effects of pH and chemical structure.

The present study demonstrates a spectroscopic study on the interaction of two phenazinium-based cationic photosensitizers, namely, phenosafranin (PSF) and safranin-O (SO) with human hemoglobin (Hb) with particular emphasis on exploring the effects of pH and chemical structures of the dye molecules on the binding phenomenon. The protein (Hb) undergoes complex conformational transitions depending on the medium pH. The dye molecules exhibit a prominent fluorescence quenching following interaction with Hb under various experimental conditions (pH 3.5, 7.4, and 9.0). Our combined steady-state and time-resolved spectroscopic results provide persuasive evidence for static quenching mechanism showing that the dye:Hb interaction proceeds through ground-state complex formation. The meticulous investigations on the pH-dependence of the interaction of the dye molecules with the protein reveal a relatively strong binding of PSF as well as SO with Hb at physiological pH and alkaline pH, while the binding is weaker at acidic pH at which Hb predominantly exists as monomeric units. The binding constant for PSF:Hb interaction is K(PSF:Hb) = (1.09 ±â€¯0.06) × 106 M-1 and that of SO:Hb interaction is K(SO:Hb) = (1.34 ±â€¯0.07) × 105 M-1 at pH 7.4. However, at pH 3.5, the binding constant values are K(PSF:Hb) = (3.58 ±â€¯0.18) × 104 M-1 and K(SO:Hb) = (4.29 ±â€¯0.22) × 104 M-1 and at pH 9.0, the values are K(PSF:Hb) = (8.08 ±â€¯0.40) × 104 M-1 and K(SO:Hb) = (5.07 ±â€¯0.25) × 104 M-1. This depicts a much stronger binding interaction of the dyes with the native Hb at pH 7.4 compared to those at pH 3.5 and 9.0. Our results also unveil the effect of chemical structures of the dyes on the interaction phenomenon in the sense that the binding constant of PSF with Hb is found to be higher than that of SO at pH 7.4 and pH 9.0. The present study also focuses on exploring such important aspects of the interaction phenomena as the effect of binding of the dyes on the protein conformation by circular dichroism spectroscopy and probable binding location of the dyes within the protein scaffolds via micropolarity measurements and molecular docking simulation.

Modulation of probe-genomic DNA interaction within the confined interior of a reverse micelle: Is the bulk-like properties of water truly achieved in large reverse micelles?

The prime motivation of the present study is to explore the effect of reverse micellar confinement on the binding interaction of an anthracene-based probe 9-methyl anthroate with herring-sperm DNA. The structural modification of the genomic DNA from its native B-form to the non-native C-form and subsequently to the condensed Ψ-form as a function of the level of hydration (W0, defined as [water] / [surfactant]) of the reverse micellar core is found to reveal a remarkable regulatory role on the stability of the stacking interaction (intercalation) of the probe within the DNA helix; the interaction being progressively stabilized at higher W0. Particularly, a close perusal of the dynamical aspects of the interaction is found to be counter-intuitive to the popular notion of the properties of the confined water within the reverse micelles typically approaching bulk-like properties at sufficiently high hydration levels (W0 > 10).

Association and sequestered dissociation of an anticancer drug from liposome membrane: Role of hydrophobic hydration.

Herein, the interaction of a potent anticancer drug (Sanguinarine, SG) with dimyristoyl-l-α-phosphatidylglycerol (DMPG) liposome membrane has been investigated at physiological pH. The spectroscopic fluorescence decay results demonstrate a modification of the photophysics of SG within DMPG-encapsulated state leading to preferential stabilization of the iminium ion over the alkanolamine form. This suggests a key role of electrostatic force underlying the interaction. The complex dependence of the thermodynamic parameters on temperature yields a unique finding of a positive heat capacity change (ΔCp) indicating the signature of hydrophobic hydration. The study also demonstrates the application of ß-cyclodextrin (ßCD) as a prospective host system resulting in release of the DMPG-bound drug. A calorimetric exploration of the DMPG-ßCD interaction reveals an intrinsically complex thermodynamics of the process leading to ΔCp > 0 and thus marking the instrumental role of hydrophobic hydration which follows that the DMPG-ßCD interaction is accompanied with burial of polar molecular surfaces. A systematic investigation of the diffusion of the drug within various microheterogeneous environments by Fluorescence Correlation Spectroscopy (FCS) categorically reinforces our arguments.

Triblock-Copolymer-Assisted Mixed-Micelle Formation Results in the Refolding of Unfolded Protein.

The present work reports a new strategy for triblock-copolymer-assisted refolding of sodium dodecyl sulfate (SDS)-induced unfolded serum protein human serum albumin (HSA) by mixed-micelle formation of SDS with poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer EO20PO68EO20 (P123) under physiological conditions. The steady-state and time-resolve fluorescence results show that the unfolding of HSA induced by SDS occurs in a stepwise manner through three different phases of binding of SDS, which is followed by a saturation of interaction. Interestingly, the addition of polymeric surfactant P123 to the unfolded protein results in the recovery of ∼87% of its α-helical structure, which was lost during SDS-induced unfolding. This is further corroborated by the return of the steady-state and time-resolved fluorescence decay parameters of the intrinsic tryptophan (Trp214) residue of HSA to the initial nativelike condition. The isothermal titration calorimetry (ITC) data also substantiates that there is almost no interaction between P123 and the native state of the protein. However, the mixed-micelle formation, accompanied by substantial binding affinities, removes the bound SDS molecules from the scaffolds of the unfolded state of the protein. On the basis of our experiments, we conclude that the formation of mixed micelles between SDS and P123 plays a pivotal role in refolding the protein back to its nativelike state.

How Does Nanoconfinement within a Reverse Micelle Influence the Interaction of Phenazinium-Based Photosensitizers with DNA?

The major focus of the present work lies in exploring the influence of nanoconfinement within aerosol-OT (AOT) reverse micelles on the binding interaction of two phenazinium-based photosensitizers, namely, phenosafranin (PSF) and safranin-O (SO), with the DNA duplex. Circular dichroism and dynamic light-scattering studies reveal the condensation of DNA within the reverse micellar interior (transformation of the B-form of native DNA to ψ-form). Our results unveil a remarkable effect of the degree of hydration of the reverse micellar core on the stability of the stacking interaction (intercalation) of the drugs (PSF and SO) into DNA; increasing size of the water nanopool (that is, w 0) accompanies decreasing curvature of the DNA duplex structure with the consequent effect of increasing stabilization of the drug:DNA intercalation. The marked differences in the dynamical aspects of the interaction scenario following encapsulation within the reverse micellar core and the subsequent dependence on the size of the water nanopool are also meticulously explored. The differential degrees of steric interactions offered by the drug molecules (presence of methyl substitutions on the planar phenazinium ring in SO) are also found to affect the extent of intercalation of the drugs to DNA. In this context, it is imperative to state that the water pool of the reverse micellar core is often argued to approach bulk-like properties of water with increasing micellar size (typically w 0 ≥ 10), so that deviation from the bulk water properties is likely to be minimized in large reverse micelles (w 0 ≥ 10). On the contrary, our results (particularly quantitative elucidation of micropolarity and dynamical aspects of the interaction) explicitly demonstrate that the bulk-like behavior of the nanoconfined water is not truly achieved even in large reverse micelles.

Interaction of phenazinium-based photosensitizers with the 'N' and 'B' isoforms of human serum albumin: Effect of methyl substitution.

The present work is focused on exploring the interaction of two phenazinium-based biological photosensitizers, phenosafranin (PSF) and safranin-O (SO), with human serum albumin (HSA), with particular emphasis on the physiologically significant NB conformational transition of the protein on the dye:HSA interaction. In addition, the presence of methyl substitution on the planar phenazinium ring in SO paves way for looking into the effect of simple chemical manipulation (that is, methyl substitution on the dye nucleus) on the dye:protein interaction behavior as a function of various (pH-induced) isoforms of HSA. Our results reveal a significantly stronger binding interaction of SO with the B isoform of HSA (at pH9.0) compared to that with the N isoform (at pH7.4). On the contrary, the PSF:HSA interaction is found to be reasonably insensitive to the aforesaid conformational transition of HSA. However, the probable binding location of both the dye molecules (PSF and SO) is found to be within the protein scaffolds (domain IB). This is further quantified from the modulation of fluorescence decay behavior of the dyes within the protein scaffolds. It is important to note that the rotational relaxation behavior of the protein-bound dyes reveals an unusual 'dip-rise-dip', an observation not reported earlier. Such unusual anisotropy decay is meticulously analyzed by an associated (or multicomponent) exponential decay model which emphasizes on the fractional contributions from differential classes of fluorophore populations characterized by the fast (due to unbound or solvent exposed part of the fluorophore) and slow (due to embedded or bound part) motions, in combination with their different local mobilities. Furthermore, the translational diffusion of the dye molecules in the presence of the protein in different isoforms (N-form or B-form) at a single molecule level is also measured by Fluorescence Correlation Spectroscopy (FCS).

Differential interaction behaviors of an alkaloid drug berberine with various bile salts.

The present study reports a meticulous characterization of the interaction of a potent anti-cancer drug, berberine chloride (BR) with a series of bile salt aggregates having varying hydrophobicity (sodium deoxycholate (NaDC), sodium cholate (NaC), and sodium taurocholate (NaTC)). The absorption spectrum of BR reveals a complex profile comprised of four distinct peaks. Analysis of the modulations of the absorption spectral properties of BR following interaction with the bile salts raising deeper questions unveils a critical insight into the mode of interaction of the cationic drug (BR) with the bile salts; a greater degree of perturbation of the microenvironment of the isoquinolinic part of BR compared to the benzenoid part. The remarkable modulation of the fluorescence profile of BR with added bile salts provides a sensitive indicator for monitoring the interaction scenario. However, an intriguing observation in this context reveals differential fluorescence behavior of BR in various bile salt aggregates, that is, similar observations in NaDC and NaC (which are legitimately interpreted according to the 'two-step association model') in comparison to NaTC. Such contrasting behavior of BR in NaTC aggregates has been rationalized on the basis of the possibility of formation of dye aggregate facilitated because of the proximity of the cationic drug molecules to the anionic headgroup of NaTC bile salt. Surprisingly, our spectroscopic results evidence for binding location of the drug at the interfacial region in all the bile salt aggregates. To this end, the time-resolved fluorescence decay behavior of the drug within various bile salt aggregates has been meticulously studied. The fluorescence decay results are found to be highly sensitive to the structure and size of the bile salt aggregates eventually leading to characterization of the interaction of the drug with the bile salts in excellent corroboration with the steady-state data. Furthermore, the time-resolved fluorescence anisotropy decay measurements yielded insight into the modulation of rotational dynamical behavior of the drug within the bile salt aggregates.

Stepwise unfolding of Ribonuclease A by a biosurfactant.

The interaction of Ribonuclease A (RNase A) with the bile salt, sodium deoxycholate (NaDC) is meticulously investigated using various spectroscopic techniques. The binding isotherm constructed from the modulation of intrinsic tyrosine fluorescence of the protein following interaction with NaDC conspicuously reveals an intrinsically complex stepwise interaction process which proceeds through the formation of distinct conformational states of the protein. The conformational transitions are found to occur at c1=2.2mM, and c2=7.2mM of NaDC. These results are subsequently corroborated from the studies of excited-state relaxation dynamics of the intrinsic tyrosine residues of RNase A, and the modulations in fluorescence behavior of an extrinsic probe (ANS) bound to the protein. The far-UV circular dichroism spectral analyses unveil only nominal influence on the secondary structural element of the protein up to [NaDC]=c1=2.2mM, which is then followed by marked disruption of the secondary structure of RNase A following further addition of NaDC. This clearly accounts for differential interaction behaviors of RNase A with the monomeric and micellar forms of NaDC (CMC of NaDC in aqueous buffer is estimated to be ∼3.0mM). In Region-I (up to c1=2.2mM), the protein:surfactant interaction is argued to be predominantly governed by electrostatic/ionic interaction force. Subsequently, in Region-II (up to c2=7.2mM) the RNase A:NaDC interaction accompanies major denaturation of the protein (∼17% loss of the secondary structure of RNase A at c2=7.2mM) resulting in significant exposure of hydrophobic surfaces of the protein. However, the tertiary structure of the protein remains essentially unperturbed within the concentration regime of NaDC under study.

Binding of ciprofloxacin to bovine serum albumin: Photophysical and thermodynamic aspects.

The present work reveals the study of interaction of a promising chemotherapeutic drug ciprofloxacin (CFX) with a model transport protein bovine serum albumin (BSA). The occurrence of the drug-protein interaction is found to result in significant modulations of the fluorescence excitation and emission spectroscopic properties of CFX following interaction with BSA. However, the major focus of the study underlies a critical insight into the quantitation of the drug-protein interaction phenomenon. To this end, we have exploited the isothermal titration calorimetric (ITC) technique to quantify the affinity constant (Ka) and stoichiometry (n) of the CFX-BSA interaction with simultaneous revelation of the accompanying thermodynamics of the interaction process. In this context, our discussion also sheds light on the lacuna surrounding various experimental methodologies for evaluation of drug-protein binding parameters. Our endeavor also extends to elucidation of the kinetic parameters and energy of activation (Ea) of the CFX-BSA interaction. The present study also delineates the modulation of the dynamical aspects of CFX following interaction with BSA. The rotational relaxation dynamics of the protein-bound drug reveals the not-so-common "dip-rise-dip" anisotropy decay. Furthermore, the effect of drug binding on the native protein conformation has been evaluated from circular dichroism (CD) spectroscopy which reveals partial rupture of the protein secondary structure.

Interaction of Bile Salts with ß-Cyclodextrins Reveals Nonclassical Hydrophobic Effect and Enthalpy-Entropy Compensation.

Herein, we present an endeavor toward exploring the lacuna underlying the host:guest chemistry of inclusion complex formation between bile salt(s) and ß-cyclodextrin(s) (ßCDs). An extensive thermodynamic investigation based on isothermal titration calorimetry (ITC) demonstrates a dominant contribution from exothermic enthalpy change (ΔH < 0) accompanying the phenomenon of inclusion complex formation, along with a relatively smaller contribution to total free energy change from the entropic component. However, the negative heat capacity change (ΔCp < 0) displays the hallmark for a pivotal role of hydrophobic effect underlying the interaction. Contrary to the classical hydrophobic effect, such apparently paradoxical thermodynamic signature has been adequately described under the notion of "nonclassical hydrophobic effect". On the basis of our results, the displacement of disordered water from hydrophobic binding sites has been argued to mark the enthalpic signature and the key role of such interaction forces is further corroborated from enthalpy-entropy compensation behavior showing indication for almost complete compensation. To this end, we have quantified the interaction of two bile salt molecules (namely, sodium deoxycholate and sodium glycocholate) with a series of varying chemical substituents on the host counterpart, namely, ßCD, (2-hydroxypropyl)-ßCD, and methyl ßCD.

Contrasting Effects of Salt and Temperature on Niosome-Bound Norharmane: Direct Evidence for Positive Heat Capacity Change in the Niosome:ß-Cyclodextrin Interaction.

The modulation of the prototropic equilibrium of a cancer cell photosensitizer, norharmane (NHM), within a niosome microheterogeneous environment has been investigated. The contrasting effects of temperature and extrinsically added salt on the photophysics of niosome-bound drug have been meticulously explored from steady-state and time-resolved spectroscopic techniques. The cation ⇌ neutral prototropic equilibrium of NHM is found to be preferentially favored toward the neutral species with increasing salt concentration, and the results are rationalized on the basis of water penetration to the hydration layer of niosome. The effects are typically reversed with temperature. The differential rotational relaxation behavior of NHM under various conditions has also been addressed from fluorescence anisotropy decay. Further, the study delineates the application of ß-cyclodextrin (ßCD) as a potential host system, leading to drug sequestration from the niosome-encapsulated state. To this end, a detailed investigation of the thermodynamics of the niosome:ßCD interaction has been undertaken by isothermal titration calorimetry (ITC) to unravel the notable dependence of the thermodynamic parameters on temperature. Consequently, a critical analysis of the variation of the enthalpy change (ΔH) of the process with temperature leads to the unique observation of a positive heat capacity change (ΔCp) marking the hallmark of hydrophobic hydration.