Blue Shift Of Emission Maximum Research Articles

Binding of a Drug (Colchicine) to L-Asparaginase Enzyme Using Multispectroscopic, Thermodynamics, and Simulation Studies: Possible Implication in Acute Lymphoblastic Leukemia Treatment.

The research aims to elucidate how drug interactions affect the activity of L-asparaginase (L-ASNase), an essential enzyme in cancer treatment, especially for acute lymphoblastic leukemia (ALL). Understanding these interactions is crucial for optimizing treatment effectiveness and reducing adverse effects. This study explores the intricate molecular interactions and structural dynamics of L-ASNase upon binding with colchicine. Fluorescence quenching experiments were conducted at various temperatures (298, 303, and 310 K), revealing notable interactions between L-ASNase and colchicine. These interactions were characterized by a reduction in fluorescence intensity and a blue shift in emission maxima. Additional analyses, including the determination of Stern-Volmer quenching constants (KSV), bimolecular quenching rate constants (kq), and thermodynamic parameters, indicated a static quenching mechanism with moderate binding affinities (Ka: 1.40-2.71 × 104 M-1) across different temperatures. Thermodynamic study suggested positive enthalpy and entropy changes (ΔH° = -10.26 kcal mol-1; ΔS° = -14.19 cal mol-1 K-1), suggesting a spontaneous reaction with negative ΔG° values (-5.86 to -6.03 kcal mol-1). FRET measurements supported optimal distances (r and Ro) for FRET occurrence, reinforcing the static quenching mechanism. Molecular docking further supported these findings, revealing a 1:1 stoichiometric binding ratio for L-ASNase:colchicine and elucidating specific binding orientations and interactions critical for complex stability. Subsequent molecular dynamics simulations spanning 100 ns underscored the stability of the L-ASNase-colchicine complex, with minimal deviations observed in key structural parameters such as RMSD, RMSF, Rg, and SASA. Additionally, spectroscopic analyses, including circular dichroism (CD), synchronous fluorescence, and 3D fluorescence provided insights into the conformational changes and alterations in the microenvironment of aromatic amino acid residues in L-ASNase upon colchicine binding. Moreover, L-ASNase activity was slightly reduced by 25% in the presence of colchicine. This comprehensive investigation sheds light on the molecular intricacies of the L-ASNase-colchicine complex, advancing our understanding of drug-target interactions and offering potential avenues for therapeutic applications.

Construction of an ultrasensitive hypochlorite fluorescent probe based on a novel chromophore fragmentation strategy

The excess intake of hypochlorite (ClO−) in the human body and excessive emission in wastewater can pose serious health risks and environmental pollution. Therefore, it is vital to develop an efficient method for detecting ClO− in foodstuff and water. In this work, we reported a lactam-fused aza-BODIPY (LAB1) for ultrasensitive ClO− detection based on novel chromophore fragmentation strategy for the first time. It exhibited distinct advantages of good selectivity, high sensitivity, fast response (60 s) and low detection limit of 9.65 nM. The sensing mechanism revealed LAB1 core was fragmented by ClO−, leading to significantly colorimetric and ratiometric fluorescence changes. As a result, 320 nm blue-shift in absorption maximum (from green to colorless) and 340 nm blue-shift in emission maximum (from dark to bright blue) allowed for visual detection by the naked eyes. The potential applicability of LAB1 for ClO− detection with satisfactory recovery in the real samples (tap water and lake water) and vegetables (spinach, lettuce, Chinese cabbage, bean sprouts), and fluorescence “turn on” imaging in mung bean sprout were demonstrated. In addition, LAB1-loaded test paper was suitable for semi-quantitative scanning ClO− analysis by using color recognition APP in a smartphone. Therefore, this work highlights a promising approach based on novel sensing mechanism for monitoring ClO− in food samples.

Exploring the synergistic effect of aggregation and hydrogen bonding: a fluorescent probe for dual sensing of phytic acid and uric acid.

We synthesized an unoxidized bis-indolyl methane (BIM) derivative (probe 1) comprising of tetraphenylethylene (TPE) as the signalling moiety. The amphiphilic probe could form self-assembled nanoscopic aggregates in the aqueous medium. The emission of 1 in non-polar solvents originates from the LE state, while in polar solvents, it is dominated by TICT. Moreover, probe 1 exhibited a 'turn-on' fluorescence response for both uric acid (with a blue shift in emission maxima) and phytic acid (with a red shift in emission maxima). Therefore, the present system provides an exceptional opportunity to distinguish between phytic acid and uric acid by considering two different emission channels. Mechanistic investigations revealed that both H-bonding and electrostatic interactions between the probe and analytes could effectively cause restricted intramolecular rotations, leading to a turn-on response. Additionally, in the case of phytic acid, larger aggregates were observed with prominent CT characteristics. The change in the extent of charge transfer interaction in the formed adducts resulted in distinct fluorescence responses with phytic acid and uric acid. Furthermore, we explored the applicability of the present system in the screening of real-life samples, such as uric acid in urine samples and phytic acid in grains. The LOD for phytic acid and uric acid was found to be ∼5.48 nM and 10.4 nM, respectively. The quantitative nature of the system was confirmed, showing promising results in terms of recovery values (between 95.6% and 104.2%) and detection limits. Additionally, we also employed handy paper strips for the on-site monitoring of phytic acid and uric acid, thereby eliminating the need for complex instrumentation or trained technicians.

Synthesis of Galactoacrylamide and Study of Stimuli‐Responsive Fluorescent Hierarchical Self‐Assembly‐Promoted Specific Interactions with Proteins

AbstractA new sugar based acrylamide, 6‐N‐acrylamido‐D‐galactopyranose (Gal‐acryl) was synthesized from D‐galactose in 6 steps. A solution of Gal‐acryl at basic pH exhibited concentration dependent fractal type assembly formation upon solvent evaporation as revealed by the microscopic images. The fractal dimensions calculated point towards Diffusion Limited Aggregation mode of aggregate formation. The critical aggregation concentration (CAC) of Gal‐acryl in water was found to be 49.3 μM. The increase in fluorescence intensity accompanied by blue shift in emission maxima of coumarin 153 suggests increase in non‐polar environment with Gal‐acryl concentration. The I1/I3ratio of pyrene emission confirmed more non‐polar microenvironment at acidic pH and vice versa at basic pH. Self‐assembly allows Gal‐acryl to exhibit specificity to lectin RCA120 as is demonstrated by fluorescence study further supported by turbidimetric assay. The fluorescence quenching studies with FITC‐RCA120 afforded the Ka value as 6.34×105 M−1, which is much higher than Ka values observed for small galactose derivatives. The present study may help in designing different fractal like glycomaterials with potential applications in drug delivery, bio‐sensing, tissue engineering, and as ligands to asialoglycoprotein receptors that are exclusively expressed by liver cells.

3-Hydroxy-2-naphthoic hydrazide as a probe for fluorescent detection of cyanide and aluminium ions in organic and aquo-organic media and its application in food and pharmaceutical samples

The commercially available fluorophore, 3-hydroxy-2-naphthoic hydrazide (RS2), has rationally been selected for the study, which displays a rapid fluorescent response and high sensitivity for CN− and Al(III) ions in neat DMSO and H2O-DMSO (1:1 v/v) media. The addition of CN− to RS2 triggers an enhancement in fluorescence at 505 nm (green fluorescence), while the addition of Al(III) increases the fluorescence of the probe with a blue-shift of emission maximum by 25 nm (bluish-green fluorescence). The probe's action was investigated by 1H NMR titrations that indicate deprotonation of OH and NH moieties by these ions. 27Al NMR of RS2-Al(III) complex suggests an octahedral geometry for the complex. The sensitivity of the fluorescent-based assays in aq. DMSO medium, 0.8 µM for CN− and 1.9 µM for Al(III) ions are far below the limits in the World Health Organization guidelines for drinking water. RS2 detects Al(III) by the chelation-enhanced fluorescence (CHEF) mechanism. Besides, RS2 was successfully applied to detect CN− and Al(III) ions in food materials and pharmaceutical samples, respectively.

An ESIPT blocked highly ICT based molecular probe to sense Zn (II) ion through turn on optical response: Experimental and theoretical studies

A simple and low cost optical sensor (L) is synthesized by the condensation reaction and characterized systematically. Among the various tested metal ions, L selectively interacts with Zn2+ ion by inducing a 17-fold high fluorescence intensity with a 60 nm blue shift of emission maxima, changing colorless solution to intense yellow solution in 20 % aqueous acetonitrile. A mechanism for selective turn on optical response of L for Zn2+ was studied using UV–vis absorption and Fluorescence emission and is correlated with theoretical results. We propose that, a dual mechanism (ESIPT/ICT) is responsible for selective sensing of Zn2+ in 20 % aqueous acetonitrile. The Job’s plot, B-H plots and ESI-MS studies confirm the 1:1 stoichiometry between host-guest interaction. A reversible optical probe with nanomolar level detection limit is non-toxic and it could render pH dependent fluorescence measurements in a live physiological environment. The practical application of L for Zn2+ was explored through bio-imaging experiment in HeLa cells.

Supramolecular-Assisted RNA-Templated Fluorescing Colloidal CdSe QDs Organized in Porous Morphology in the Presence of 1,3-Diaminopropane: Study of Their Multifunctional Behavior

The synthesis of colloidal CdSe nanostructures mediated by ribonucleic acid (RNA) in the presence of 1,3-diaminopropane (DAP) and excess Cd2+ produces CdSe QDs (average diameter, 4.5 nm) forming a chainlike morphology (SP1-DAP) with fairly high fluorescence efficiency and average lifetime (⟨τ⟩): 36% and 121 ns (λflu,sh = 650 nm), respectively. The addition of DAP induces its transformation to porous morphology in the process of self-assembly (SP1A-DAP), as evidenced by atomic force microscopy and transmission electron microscopy, and increase in rotational time constant from 14.7 to 31.5 ns. These changes are associated with a blue shift in emission maxima from 545 to 530 nm and an increase in ⟨τ⟩ from 102 to 111 ns (λmax,flu = 545 nm). For SP1A-DAP, the development of distinct bands (cm–1) in IR analysis (1240 (s) and 1183 (sh)), blue shift in positive peak at 271.1 nm in circular dichroism spectra, and changes in 1H and 31P NMR spectra indicating RNA configuration change from C2′-endo to C3′-endo, clear...

Blue luminescence from ZnGa2O4: Effect of lattice distortion and particle size

Dependence of structural and luminescence properties of nano-crystalline ZnGa2O4 phosphors on the synthesis conditions such as concentration of precursor metal ions and nature of solvents were investigated in detail. Based on XRD and luminescence studies carried out on samples prepared with high and low concentrations of precursor metal ions in ethylene glycol medium, it is established that distortion of structural units along with defects/traps in the lattice, and not the particle size, is responsible for the observed blue shift in emission maxima with decrease in precursor concentrations. Unlike this, a red shift is observed when the synthesis is carried out under identical conditions in water medium. This has been explained based on difference in particle size distributions as confirmed by TEM studies.

Binding of the alkaloid coralyne to parallel G-quadruplex DNA [d(TTGGGGT)]4 studied by multi-spectroscopic techniques

Binding of coralyne to tetra-molecular parallel G-quadruplex DNA [d(TTGGGGT)]4 was evaluated for the first time using biophysical techniques. Absorbance titrations show hypo/hyper-chromism accompanied by 12nm red shift with binding constant Kb=0.2–4.0×106M−1. Binding induces a negative circular dichroism band of coralyne at 315nm. Quenching of fluorescence (~64%) along with 10nm blue shift in emission maxima indicates proximity of coralyne to guanine bases. Job plot indicates existence of multiple complexes. The observed two fluorescence life times, 6 and 12ns, with relative abundance 33% and 63%, respectively suggest two binding sites/conformations in complex. 1D 1H NMR spectra reveal significant broadening and upfield shift of G3NH, G6NH and G3H8 proton signals upon binding. The NOESY spectra reveal sequence specific non-intercalative interaction of coralyne in monomeric form at two sites close to A3/G3 and G6 bases of [d(TTGGGGT)]4 and [d(TTAGGGT)]4, which has implications in anti-cancer drug action.

Probing microenvironment of micelle and albumin using diethyl 6-(dimethylamino)naphthalene-2,3-dicarboxylate: An electroneutral solvatochromic fluorescent probe

Synthesis, single-crystal X-ray characterization, and spectroscopic investigation of small, non-charged diethyl 6-(dimethylamino)naphthalene-2,3-dicarboxylate (DMNDC) by UV–Visible, steady-state, and time-resolved fluorescence reveal a series of interesting photophysical properties originating from the intrinsic intramolecular charge transfer (ICT) state, leading to diverse applications. Stokes shift, lifetime, and emission maxima of DMNDC show a very good correlation with ET(30) solvent polarity scale for a series of different polarity solvents, confirms that it has very good environment sensitivity. Furthermore, this dye has been found to be an exceptionally suitable probe for determining Critical Micelle Concentration (CMC) and probing self-organization processes of five different type of surfactant with structural diversity. A 20–60nm blue shift in emission maxima accompanied by a large fluorescence lifetime enhancement (ca. 23ns) was observed upon relocation of DMNDC into a hydrophobic microenvironment. Along with this, the small size, electroneutrality, pH stability, and excellent solvatochromic fluorescent properties are employed for deciphering the number of hydrophobic binding pockets with strong affinity and their local microenvironment present in Bovine Serum Albumin (BSA).

Contrasting Solid-State Fluorescence of Diynes with Small and Large Aryl Substituents: Crystal Packing Dependence and Stimuli-Responsive Fluorescence Switching.

There has been a significant current interest in solid state luminescence of organic molecules and their stimuli responsive fluorescence switching behavior. Although small organic derivatives with olefinic, acetylenic, phenylenevinylenic, phenyleneethynylenic spacers are widely documented as solid state emitters in the literature, the solid state photophysics of organic derivatives with "butadiyne" spacer still remains unexplored. We provide detailed investigation on the solid state fluorescence properties of a series of butadiynyl fluorophores. Replacement of a phenyl ring, which is at periphery of the butadiyne bridge, with a large moiety such as pyrenyl group furnishes contrasting emissions in the solid state. While the butadiyne bridged phenyl derivatives show a blue shift of emission maxima in the solid powder with respect to monomer spectra in solution state, the butadiyne bridged pyrenyl derivatives exhibit a red shift in the solid state. The blue shift of the emission maxima of the butadiyne bridged phenyl derivatives in the solid powder is attributed to allowed excitonic transition in aggregates with nearly parallel transition dipoles. On the other hand, formation of pyrenyl excimer accounts for the red shift of the butadiyne bridged pyrenyl derivatives in the solid powder. In addition to that, the solid state fluorescence of the pyrenyl analogues is reversibly switched between two aggregate forms through external heating and rubbing stimuli.

Investigation on luminescence enhancement and decay characteristics of long afterglow nanophosphors for dark-vision display applications

Long afterglow SrAl2O4:Eu2+,Dy3+ nanophosphors were synthesized via a facile but effectual auto-combustion technique followed by post-annealing treatment at elevated temperatures. The influence of various fuels during synthesis and thereafter improvement in the luminescence decay characteristics under various illuminant irradiations of long afterglow nanophosphors have been reported. Extensive studies on structural, morphological and luminescent properties of the as-synthesized afterglow nanophosphors have been presented. Powder X-ray diffraction studies confirm the presence of high-purity, single-phase monoclinic nanophosphors. HRTEM investigations confirm the formation of nanophosphors of particle size less than 50nm. Photoluminescence emission is attributed to the characteristic d–f transition (4f65d1→4f7) of Eu2+ ions and was positioned at 512nm. As-synthesized nanophosphors exhibit considerable confinement effects resulting into blue shift in emission maxima as compared to their bulk counterparts. The mechanism underlined for long afterglow has been discussed using trapping–detrapping model. The nanophosphor being multifunctional finds many interesting applications including dark-vision display, energy storage, fingerprint detection, in vivo and in vitro biological staining, etc.

Optimising conditions for the growth of nanocrystalline ZnS thin films from acidic chemical baths

The growth of nanocrystalline zinc sulfide thin films onto glass substrates by chemical bath deposition has been optimized at acidic pH. Powder X-ray diffraction (p-XRD) confirms the deposition of sphalerite, the cubic phase of ZnS. The crystallite size calculated by Scherrer equation was found to be 4.0nm. Scanning Electron Microscopy (SEM) show clusters of spherical nanoparticles uniformly distributed over the surface of the glass substrates. Energy Dispersive X-ray (EDX) analysis of the deposited thin films show the zinc to sulfur ratio close to 1:1. The observed band gap (3.78eV) of the deposited thin films is higher than that reported for cubic phase of bulk ZnS (3.54eV) as expected due to nano-size crystallites. Binding energies calculated by X-ray Photoelectron Spectroscopy (XPS) confirm the material as ZnS and the photoluminescence measurements show the blue shift in emission maximum.

Elucidating the mode of action of urea on mammalian serum albumins and protective effect of sodium dodecyl sulfate

The effect of sodium dodecyl sulfate (SDS) on human, bovine, porcine, rabbit and sheep serum albumins were investigated at pH 3.5 by using various spectroscopic techniques like circular dichroism (CD), intrinsic fluorescence and dynamic light scattering (DLS). In the presence of 4.0mM SDS the secondary structure of all the albumins were not affected as measured by CD but fluorescence spectra revealed 8.0nm blue shift in emission maxima. We further checked the stability of albumins in the absence and presence of 4.0mM SDS by urea and temperature at pH 3.5. In the absence of SDS, urea starts unfolding both secondary as well as tertiary structural elements of the all the albumins at ∼2.0M urea but in the presence of 4.0mM SDS, urea was unable to unfold even up to 9.0M. The albumins were thermally less stable at pH 3.5 with decrease in Tm but in the presence of 4.0mM SDS, the Tm was increased. From this study, it was concluded that SDS is showing a protective effect against urea as well as thermal denaturation of albumins. This behavior may be due to electrostatic as well as the hydrophobic interaction of SDS with albumins. Further, we have proposed the mechanism of action of urea. It was found that urea interacted with proteins directly when proteins are in charged form. Indirect interaction may be taking place when the environment is more hydrophobic.

Salicylaldehyde Phenylhydrazone: A New Highly Selective Fluorescent Lead (II) Probe

The fluorescence intensity of salicylaldehyde phenylhydrazone (L), in 1:1 (v/v) CH3OH:H2O was enhanced by ca. 100 times with a blue shift in emission maximum, on interaction with Pb(2+) ion. No enhancement in fluorescent intensity of L was observed on interaction with metal ions - Na(+), K(+), Ca(2+), Cu(2+), Ni(2+), Zn(2+), Cd(2+) and Hg(2+). This signal transduction was found to occur via photoinduced electron transfer (PET) mechanism. A 1:1 complexation between Pb(2+) and L with log β = 7.86 has been proved from fluorescent and UV/Visible spectroscopic data. The detection limit of Pb(2+) was calculated to be 6.3 × 10(-7) M.

Ligand binding induces a sharp decrease in hydrophobicity of folate binding protein assessed by 1-anilinonaphthalene-8-sulphonate which suppresses self-association of the hydrophobic apo-protein

High affinity folate binding protein (FBP) regulates as a soluble protein and as a cellular receptor intracellular trafficking of folic acid, a vitamin of great importance to cell growth and division. We addressed two issues of potential importance to the biological function of FBP, a possible decrease of the surface hydrophobicity associated with the ligand-induced conformation change of FBP, and protein-inter-protein interactions involved in self-association of hydrophobic apo-FBP.The extrinsic fluorescent apolar dye 1-anilinonaphthalene-8-sulphonate (ANS) exhibited enhanced fluorescence intensity and a blueshift of emission maximum from 510–520 to 460–470nm upon addition of apo-FBP indicating binding to a strongly hydrophobic environment. Neither enhancement of fluorescence nor blueshift of ANS emission maximum occurred when folate-ligated holo-FBP replaced apo-FBP. The drastic decrease in surface hydrophobicity of holo-FBP could have bearings on the biological function of FBP since changes in surface hydrophobicity have critical effects on the biological function of receptors and transport proteins.ANS interacts with exposed hydrophobic surfaces on proteins and may thereby block and prevent aggregation of proteins (chaperone-like effect). Hence, hydrophobic interactions seemed to participate in the concentration-dependent self-association of apo-FBP which was suppressed by high ANS concentrations in light scatter measurements

Novel Benzanthrone Aminoderivatives for Membrane Studies

The potential of novel benzanthrone aminoderivatives to trace the changes in physicochemical properties of lipid bilayer has been evaluated. Binding of the dyes to the lipid bilayers composed of zwitterionic phospholipid phosphatidylcholine (PC) and its mixtures with anionic phospholipid cardiolipin (CL) and cholesterol (Chol) was followed by significant quantum yield increase with small blue shift of emission maximum. Analysis of partition coefficients of the dyes under study showed that all aminobenzanthrones possess high lipid-associating ability. The dyes A8 and AM2 proved to be sensitive to the variations in membrane chemical composition responding to the changes in bilayer hydration induced by CL and Chol.

A novel approach to electrospinning of pristine and aligned MEH-PPV using binary solvents

Pristine and aligned electrospun fibers of poly[2-methoxy-5-(2′-ethylhexyloxy)-1,4-phenylenevinylene] (MEH-PPV) have been fabricated by electrospinning. The key to success was using a binary solvent system, in which a poor solvent was introduced to increase interchain interactions of MEH-PPV. At the same time, the poor solvent should have a low surface tension and high conductivity. The emission spectra of pristine MEH-PPV in electrospun fibers, spin coating film and dilute solution were acquired and analyzed. The emission anisotropy of aligned electrospun fibers was investigated by polarized emission measurement. For pristine MEH-PPV fibers, the emission anisotropy is 0.47 at emission peak, indicating alignment of MEH-PPV chains along the electrospun fiber axis. For MEH-PPV/poly (e-caprolactone) (PCL) electrospun fibers, the interchain energy transfer in MEH-PPV was significantly depressed. As a result, MEH-PPV/PCL electrospun fibers exhibited a strong blue shift of emission maximum and increased emission anisotropy of 0.66 compared with pristine MEH-PPV electrospun fibers. The ease of electrospinning of pristine MEH-PPV provides new opportunities for its application in unique optoelectronic devices.

Effect of Sr 2+-doping on structure and luminescence properties of BaAl 2Si 2O 8:Eu 2+ phosphors

Sr 2+ doped BaAl 2Si 2O 8:Eu 2+ phosphor was synthesized by chemical co-precipitation method. With the increase of Sr 2+ concentration, the phase structure of (Ba 0.965 − x Sr xEu 0.035)Al 2Si 2O 8 changes from hexagonal phase to monoclinic phase owing to large activation energy in SrAl 2Si 2O 8 system. (Ba 0.965 − x Sr xEu 0.035)Al 2Si 2O 8 phosphor exhibits a broad blue band peaking at 425 nm due to the 4f 65d–4f 7( 8S 7/2) transition of Eu 2+ ions. The emission intensity increases, accompanied by the blue shift of emission maximum from 459 to 417 nm with the Sr 2+ doping concentration increasing. The optimal concentration of Sr 2+ ion is 40%, and the phosphor shows high color stability in CIE chromaticity diagram. The result indicates that Sr 2+ doped phosphor not only can enhance the relative intensity but also can adjust the chromaticity coordinate.

Partitioning and localization of environment-sensitive 2-(2'-pyridyl)- and 2-(2'-pyrimidyl)-indoles in lipid membranes: a joint refinement using fluorescence measurements and molecular dynamics simulations.

Fluorescence of environment-sensitive dyes is widely applied to monitor local structure and solvation dynamics of biomolecules. It has been shown that, in comparison with a parent indole fluorophore, fluorescence of 2-(2'-pyridyl)-5-methylindole (5M-PyIn-0) and 2-[2'-(4',6'-dimethylpyrimidyl)]-indole (DMPmIn-0) is remarkably sensitive to hydrogen bonding with protic partners. Strong fluorescence, observed for these compounds in nonpolar and polar aprotic solvents, is efficiently quenched in aqueous solution. This study demonstrates that 5M-PyIn-0 and DMPmIn-0, which are almost nonemitting in aqueous solution, become highly fluorescent upon titrating with phospholipid vesicles. The fluorescence enhancement is accompanied by a significant blue shift of emission maximum. The Gibbs free energy of membrane partitioning, measured by the increase in the steady-state fluorescence intensities during transfer from an aqueous environment to a lipid bilayer, is very favorable for both compounds, being in a range from -7.1 to -8.0 kcal/mol and depending only slightly on lipid composition of the membrane. The fluorescence enhancement upon membrane partitioning is indicative of the loss of the specific hydrogen-bonding interactions between the excited fluorophore and water molecules, causing efficient fluorescence quenching in bulk water. This conclusion is supported by atomistic molecular dynamics (MD) simulations, demonstrating that both 5M-PyIn-0 and DMPmIn-0 bind rapidly and partition deeply into a lipid bilayer. MD simulations also show a rapid, nanosecond-scale decrease in the probability of solute-solvent hydrogen bonding during passive diffusion of the probe molecules from bulk water into a lipid bilayer. At equilibrium conditions, both 5M-PyIn-0 and DMPmIn-0 prefer deep localization within the hydrophobic, water-free region of the bilayer. A free energy profile of penetration across a bilayer estimated using MD umbrella sampling shows that both indole derivatives favor residence in a rather wide potential energy well located 10-15 Å from the bilayer center.