Charged Head Groups Research Articles

Atomistic Mechanism of Lipid Membrane Binding for Blood Coagulation Factor VIII with Molecular Dynamics Simulations on a Microsecond Time Scale.

During the blood coagulation cascade, coagulation factor VIII (FVIII) is activated by thrombin to form activated factor VIII (FVIIIa). FVIIIa associates with platelet surfaces at the site of vascular damage to form an intrinsic tenase complex with activated factor IX. A working model for FVIII membrane binding involves the association of positively charged FVIII residues with negatively charged lipid headgroups and the burial of hydrophobic residues into the membrane interior. Currently, the atomic details of the FVIII lipid binding interactions and membrane orientation are lacking. This study reports residue-specific FVIII C domain interactions with 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (DOPS) in atomistic detail. Contact maps between residues in the C domains with different lipid moieties support prior structural data describing how the C domains associate with membranes through electrostatic and hydrophobic interactions. Solvent-accessible surface area analysis quantified the extent to which residues in the C1 and C2 domains bury into the membrane. Calculations of the potential energy between the C domains and DOPC and DOPS revealed an FVIII membrane-binding orientation that agrees with previous experimental data. This study expands our knowledge of the structural basis of FVIII membrane association, which may be critical for the development of next-generation FVIII replacement constructs with improved activity.

Interactions between Per- and Polyfluoroalkyl Substances (PFAS) at the Water-Air Interface.

Per- and polyfluoroalkyl substances (PFAS)─so-called "forever chemicals"─contaminate the drinking water of about 100 million people in the U.S. alone and are inefficiently removed by standard treatment techniques. A key property of these compounds that underlies their fate and transport and the efficacy of several promising remediation approaches is that they accumulate at the water-air interface. This phenomenon remains incompletely understood, particularly under conditions relevant to natural and treatment systems where water-air interfaces often carry significant loads of other organic contaminants or natural organic matter. To understand the impact of organic loading on PFAS adsorption, we carried out molecular dynamics simulations of PFAS at varying interfacial densities. We find that adsorbed PFAS form strong mutual interactions (attraction between perfluoroalkyl chains and electrostatic interactions among charged head groups) that give rise to ordered interfacial coatings. These interactions often involve near-cancellation of hydrophobic attraction and Coulomb repulsion. Our findings explain an apparent paradox whereby PFAS adsorption isotherms often suggest minimal mutual interactions while simultaneously displaying a high sensitivity to the composition and density of interfacial coatings. Consideration of the compounds present with PFAS at the interface has the potential to allow for more accurate predictions of fate and transport and the design of more efficient remediation approaches.

Hybrid Bilayer Interfaces within Reversed-Phase Chromatographic Silica Formed by Self-Assembly of Long-Chain Primary Alcohols.

Modification of silica interfaces by covalent attachment of functional ligands is a primary means of controlling the interfacial chemistry of porous silicas used in separations, environmental cleanup, and biosensing. Recently, modification of hydrophobic, n-alkyl-silane-functionalized interfaces has been achieved through self-assembly of zwitterionic phospholipids or mixed-charged surfactants to form "hybrid bilayers", producing interfaces that mimic lipid-bilayer partitioning and provide shape-selective partitioning of aromatic hydrocarbons. Charged headgroups, however, introduce electrostatic interactions that strongly influence the retention of ionizable solutes and require careful control over pH and ionic strength in the solution phase. In this work, we propose modification of C18-functionalized chromatographic silica surfaces through self-assembly of long-chain primary alcohols to form uncharged hybrid-bilayer surfaces. Hybrid bilayers formed from alcohols ranging from C12OH to C22OH are investigated with in situ confocal-Raman microscopy, and the spectra indicate that they form highly ordered n-alkane structures, with order increasing as a function of alcohol chain length. Temperature-dependent Raman spectra of C12OH-C22OH hybrid bilayers were collected to investigate their melting transitions. Multivariate curve resolution of these spectra show broad, two-component melting transitions, indicating alcohol and C18 alkyl chains melt simultaneously. These results suggest an interdigitated interfacial structure, where the hydrocarbon chains of the adsorbed alcohol extend into the underlying C18 chains, ordering both layers. Interdigitation is confirmed by a temperature-dependent study of a deuterated C16-OH bilayer, where spectrally resolved Raman bands from deuterated and protiated hydrocarbons melt together. Finally, n-alkyl alcohol bilayers were tested for protein repellency, where no protein adsorption was observed when equilibrated with ∼1 mg/mL bovine serum albumin. Bilayers C16OH in chain length are shelf stable at refrigerated temperatures for months. These results demonstrate long-chain alcohol bilayers can be utilized to control the interfacial hydrocarbon structure of C18-modified silica and have potential for use in separations, biosensing, and anti-biofouling applications.

Charge-Stabilized Nanodiscs as a New Class of Lipid Nanoparticles.

Nanoparticles have the potential to improve disease treatment and diagnosis due to their ability to incorporate drugs, alter pharmacokinetics, and enable tissue targeting. While considerable effort is placed on developing spherical lipid-based nanocarriers, recent evidence suggests that high aspect ratio lipid nanocarriers can exhibit enhanced disease site targeting and altered cellular interactions. However, the assembly of lipid-based nanoparticles into non-spherical morphologies has typically required incorporating additional agents such as synthetic polymers, proteins, lipid-polymer conjugates, or detergents. Here, charged lipid headgroups are used to generate stable discoidal lipid nanoparticles from mixed micelles, which are termed charge-stabilized nanodiscs (CNDs). The ability to generate CNDs in buffers with physiological ionic strength is restricted to lipids with more than one anionic group, whereas monovalent lipids only generate small nanoliposomal assemblies. In mice, the smaller size and anisotropic shape of CNDs promote higher accumulation in subcutaneous tumors than spherical liposomes. Further, the surface chemistry of CNDs can be modified via layer-by-layer (LbL) assembly to improve their tumor-targeting properties over state-of-the-art LbL-liposomes when tested using a metastatic model of ovarian cancer. The application of charge-mediated anisotropy in lipid-based assemblies can aid in the future design of biomaterials and cell-membrane mimetic structures.

Investigations of Crucial Factors for the Non-Covalent Functionalization of Black Phosphorus (BP) using Perylene Diimide Derivatives for the Passivation of BP Nanosheets.

The non-covalent functionalization of black phosphorus (BP) was studied with a scope of ten tailor-made perylene diimides (PDIs). A combination of UV/Vis-, fluorescence-, as well as Raman spectroscopy and atomic force microscopy was used to investigate the structural factors, which contribute to a pronounced PDI-BP interaction and thus support the protection of BP nanosheets against oxidative degradation. We were able to show, that water-soluble, amphiphilic PDIs with highly charged head groups can be used for the non-covalent functionalization of BP in aqueous media. Here, based on the hydrophobic effect, an efficient adsorption of the respective PDI molecules takes place and leads to the formation of a passivating film, yielding a considerable stabilization of the BP flakes under ambient conditions exceeding 30 days.

Modulating the behavior of ethyl cellulose-based oleogels: The impact food-grade amphiphilic small molecules on structural, mechanical, and rheological properties

This work evaluates the ability of various lipid-based amphiphilic small molecules (ASMs) to modulate the mechanical and rheological properties of oleogels principally structured by ethyl cellulose (EC). Six ASMs varying in the chemical structure of their polar headgroups were used to produce EC-ASM oleogels. Stearic acid (StAc), monoacylglycerol (MAG), sodium stearoyl lactylate (SSL), and citric acid esters of monoglycerides (CITREM) all provided a dramatic enhancement in gel strength, while lactic acid (LACTEM) and acetic acid (ACETEM) esters produced only a marginal increase. Those additives which crystallized above 20 °C displayed pronounced changes in their network organization and crystal morphology in the presence of EC. Differences in the solid/liquid phase change behavior were also observed in select samples using differential scanning calorimetry. Both the small and large amplitude oscillatory shear responses were dependent on the ASM which was dependent on the chemistry of the headgroup, crystal network organization, and ability to plasticize the polymer network. The extent of thixotropic recovery was largely dependent on the polarity of functional groups in the ASMs, but was also influenced by the formation of a secondary crystal network. In general, ASMs which formed larger, system-spanning crystal networks (MAG, StAc) produced more brittle gels, while the highly hydrophilic, charged headgroup of SSL promoted a homogeneous distribution of small crystals, resulting in a tougher material. In the absence of a crystal network, stronger polar species in the ASM headgroup produced higher gel strength and increased elasticity. Thus, both ASM chemical structure and crystallization properties strongly contribute to the functionality of the resulting combined oleogelator systems.

Insights into the dual effect of an amphiphilic additive on hydrate formation from a water structure perspective

Chemical additives play an important role in gas hydrate formation, which involves the transformation of water structure. However, the relationship between the effects of additives on water structure and hydrate formation is inconclusive, especially for amphiphilic molecules. Here, we investigate the changes in water structure caused by rhamnolipid (Rha) biosurfactants and the resulting effect on methane hydrate formation through kinetic experiments, morphological observation, and in situ Raman spectroscopy. The Gaussian peaks of strongly hydrogen-bonded and weakly hydrogen-bonded water are identified, of which the integrated intensity and position are compared. It is found that the dual effect of Rha on hydrate formation kinetics, namely nucleation inhibition and growth promotion, is attributed to the ordered arrangement of water molecules with stronger hydrogen bonding interactions induced by the charged head groups. The strongly hydrogen-bonded water inhibits nucleation but promotes growth. In addition, the adsorption of Rha on montmorillonite (MMT) eliminates the changes in water structure and the dual effect of hydrate formation kinetics, providing further evidence for the above conclusion. These insights pave the way for a deeper understanding of hydrate formation kinetics from a water structure perspective, as well as providing a scientific basis for the commercial application of surfactants.

The Role of Surfactant Head Group Charge on Binding of Hoechst 33258 with Micelles

AbstractOwing to the diverse biological application, photophysical behaviour of Hoechst 33258 (H33258) is an important topic in biophysical research. Here, we have reported the photophysical behavior of H33258 in model biomembrane mimicking micellar microenvironment. In order to understand the molecular mechanism of the interaction of H33258 with micelles, we have used cationic (Hexadecetyltrimethyl ammonium bromide (CTAB)), anionic (sodium dodecyl sulphate (SDS)), and non‐ionic (t‐octylphenoxypolyoxyethanol, Triton‐X100 (TX 100)) micelles. From the steady‐state emission study the evaluated apparent binding constant (Kb) values for the complex formation between H33258 and CTAB, SDS and TX 100 micelle were (1.45±0.42)×105, (31.95±4.89)×105 and (3.10±0.73)×105 M−1, respectively. Thus, it was found that the Kb value was greater in the case of anionic micelle than non‐ionic and cationic micelles. Such differences in Kb values were due to the electrostatic interaction and hydrogen bond formation between the probe molecule and micelles. In addition, time‐resolved fluorescence decay and time‐resolved fluorescence anisotropy studies were performed to understand the dynamical behavior of H33258 in presence of micelles. The interaction of H33258 with micelles might be useful to study the structure and dynamics of different bio‐membranes and to formulate new drug delivery system.

An insight into physicochemical properties of hexagonal lyotropic liquid crystal templates from amphiphiles for mesoporous structural administration

Hexagonal lyotropic liquid crystals (HLLC) provide an ideal template for the fabrication of nanofiltration membranes without the flux-selectivity trade-off problem. Amphiphiles, the core components of the columnar micelles in the HLLC system, play a key role in regulating the nanostructure, their effects on dimensional stability and physicochemical properties of the system however have not been explored well. In this research, we compared the dimensional stability and physicochemical properties of HLLC templates prepared from amphiphiles with various head groups and tail lengths. The amphiphiles with charged head groups (dodecyl trimethylammonium bromide, DTAB, and hexadecyl trimethylammonium bromide, CTAB) enable more monomers to be introduced and a much smaller (∼ 1/2) pore size than that with uncharged ones (polyethylene glycol hexadecyl ether, Brij56). The tail length however mainly regulates the width of water channels with CTAB being 23.27 Å and DTAB about 17.06 Å when the head groups are identical. Although the HLLC systems with CTAB and DTAB present a little bit higher phase transition temperature, the dynamic mechanical stability is much better than that with Brij56 at room temperature, facilitating the reorientation process under an electric force with the order parameter reaching to about 0.9 and the retention of the aligned nanostructure with the absence of the force at room temperature. The study enables a better understanding of the effects of amphiphiles on HLLC templates and their tunability for advanced nanofiltration membranes.

Electric charges of the lipid headgroup modulate Melittin adsorption to lipid vesicle membranes

Peptide lipid membrane interactions are modulated by factors such as peptide hydrophobicity and the electric charge of both the peptide and the membrane surface. We investigated the influence of lipid headgroup charge on the adsorption of Melittin (Mel) on the surface of artificial lipid vesicles in conditions that mimic the interaction with biological cells, such as ionic strength, pH, and peptide concentration. Taking advantage of Mel TRP residue fluorescence, we used several fluorescence techniques (FRET, fluorescence quenching, time-resolved fluorescence) to investigate Mel adsorption on the surface of the DMPC and DMPC with fractions of positively charged lipids (EPC) or negatively charged lipids (DPPG) vesicles. Our results show that the DMPC:DPPG vesicles allow a deeper localization of TRP residues in the lipid membrane, proven by the blue shift of the TRP emission spectrum and the exclusion radius evaluated from FRET. We also showed that even for positively charged surfaces Mel tends to adsorb on the lipid surface. The results were confirmed indirectly by quencher accessibility to TRP. Finally, we discuss the results in correlation to the Mel effects on biological cells

An Investigation of the Effect of pH on Micelle Formation by a Glutamic Acid-Based Biosurfactant

NMR spectroscopy, molecular modeling, and conductivity experiments were used to investigate micelle formation by the amino acid-based surfactant tridecanoic L-glutamic acid. Amino acid-based biosurfactants are green alternatives to surfactants derived from petroleum. NMR titrations were used to measure the monomeric surfactant’s primary and gamma (γ) carboxylic acid pKa values. Intramolecular hydrogen bonding within the surfactant’s headgroup caused the primary carboxylic acid to be less acidic than the corresponding functional group in free L-glutamic acid. Likewise, intermolecular hydrogen bonding caused the micellar surfactant’s γ carboxylic functional group to be less acidic than the corresponding monomer value. The binding of four positive counterions to the anionic micelles was also investigated. At pH levels below 7.0 when the surfactant headgroup charge was −1, the micelle hydrodynamic radii were larger (~30 Å) and the mole fraction of micelle-bound counterions was in the 0.4–0.7 range. In the pH range of 7.0–10.5, the micelle radii decreased with increasing pH and the mole fraction of micelle bound counterions increased. These observations were attributed to changes in the surfactant headgroup charge with pH. Above pH 10.5, the counterions deprotonated and the mole fraction of micelle-bound counterions decreased further. Finally, critical micelle concentration measurements showed that the micelles formed at lower concentrations at pH 6 when the headgroup charge was predominately −1 and at higher concentrations at pH 7 where headgroups had a mixture of −1 and −2 charges in solution.

Short-Chained Linear Scorpion Peptides: A Pool for Novel Antimicrobials.

Scorpion venom peptides are generally classified into two main groups: the disulfide bridged peptides (DBPs), which usually target membrane-associated ion channels, and the non-disulfide bridged peptides (NDBPs), a smaller group with multifunctional properties. In the past decade, these peptides have gained interest because most of them display functions that include antimicrobial, anticancer, haemolytic, and anti-inflammatory activities. Our current study focuses on the short (9-19 amino acids) antimicrobial linear scorpion peptides. Most of these peptides display a net positive charge of 1 or 2, an isoelectric point at pH 9-10, a broad range of hydrophobicity, and a Grand Average of Hydropathy (GRAVY) Value ranging between -0.05 and 1.7. These features allow these peptides to be attracted toward the negatively charged phospholipid head groups of the lipid membranes of target cells, a force driven by electrostatic interactions. This review outlines the antimicrobial potential of short-chained linear scorpion venom peptides. Additionally, short linear scorpion peptides are in general more attractive for large-scale synthesis from a manufacturing point of view. The structural and functional diversity of these peptides represents a good starting point for the development of new peptide-based therapeutics.

Critical Assessment of Micellar Surface Charge-Dependent Disaggregation and Reaggregation of a Bis-Indole Self-Aggregate: What Should Be Our Case-Dependent Choice?

"Aggregation-caused quenching" is a deep-seated mechanism and has been widely used by the researchers as the possible basis for new sensor development. Contrast to aggregation, its turn around process, disaggregation, has gained much less consideration so far. Unfortunately, study of the further scope for reaggregation of the disaggregated probe assembly in the same solution, as and when required, is still under the rare category. The central theme of the current study is focused on this aspect. For this purpose, the effects of headgroup charge (cationic, anionic, and nonionic) and polarity of the micelles on the degree of disaggregation and subsequent emission amelioration of a synthesized bis-indole derivative, 3,3'-bisindolyl(phenyl)methane (BIPM), are studied using steady-state and time-resolved spectroscopic techniques. The relative emission yield of BIPM (φf = 0.01) is significantly enhanced in the presence of cetyltrimethylammonium bromide (φf = 0.21) and polyoxyethylene (20) sorbitan monolaurate (φf = 0.24), whereas comparatively less emission enhancement is recorded within the sodium dodecyl sulfate system (φf = 0.07). In contrast, addition of an external biophilic agent, uric acid, causes requenching of the enhanced emission because of the reaggregation of the disaggregated probes. Detailed microscopic and calorimetric studies are also adopted to investigate the disaggregation-reaggregation mechanism of BIPM associations. The study will provide prior insights about the use of suitable micellar systems for the required degree of disaggregation as well as for the modulation of emission efficiency by controlled tuning of the disaggregation-reaggregation equilibrium for similar probe associations in pure aqueous medium avoiding any chemical transformation.

Effect of cationic Gemini surfactants ethonium and decamethoxin on the spectral properties, solubility and tautomerism of the curcumin

AbstractThe properties of natural polyphenol curcumin in water solutions of cationic Gemini surfactants decamethoxin and ethonium, which have similar positively charged nitrogen hydrophilic head groups but different spacer sizes and lengths of hydrophobic tails have been investigated in a wide range of concentrations by using UV–Vis spectroscopy and quantum‐chemical calculations. It was found that curcumin dissolves in the organized micellar media of these surfactants in the enol tautomeric form and its solubility increases 400 times compared to water. The binding constants of curcumin with decamethoxin and ethonium were determined by the solubility method. Significant differences in the influence of premicellar concentrations of cationic Gemini surfactants on the solubility and tautomeric transformations of curcumin were revealed. In contrast to decamethoxin, ethonium at a concentration below the CMC promotes a significant increase in the solubility of curcumin and a shift in the tautomeric equilibrium towards the formation of its keto form.

Quantitative cross-species comparison of serum albumin binding of per- and polyfluoroalkyl substances from five structural classes.

Per- and polyfluoroalkyl substances (PFAS) are a class of over 8000 chemicals, many of which are persistent, bioaccumulative, and toxic to humans, livestock, and wildlife. Serum protein binding affinity is instrumental in understanding PFAS toxicity, yet experimental binding data is limited to only a few PFAS congeners. Previously, we demonstrated the usefulness of a high-throughput, in vitro differential scanning fluorimetry assay for determination of relative binding affinities of human serum albumin for 24 PFAS congeners from 6 chemical classes. In the current study, we used this assay to comparatively examine differences in human, bovine, porcine, and rat serum albumin binding of 8 structurally informative PFAS congeners from 5 chemical classes. With the exception of the fluorotelomer alcohol 1H, 1H, 2H, 2H-perfluorooctanol (6:2 FTOH), each PFAS congener bound by human serum albumin was also bound by bovine, porcine, and rat serum albumin. The critical role of the charged functional headgroup in albumin binding was supported by the inability of albumin of each species tested to bind 6:2 FTOH. Significant interspecies differences in serum albumin binding affinities were identified for each of the bound PFAS congeners. Relative to human albumin, perfluoroalkyl carboxylic and sulfonic acids were bound with greater affinity by porcine and rat serum albumin, and the perfluoroalkyl ether acid congener bound with lower affinity to porcine and bovine serum albumin. These comparative affinity data for PFAS binding by serum albumin from human, experimental model, and livestock species reduce critical interspecies uncertainty and improve accuracy of predictive bioaccumulation and toxicity assessments for PFAS.

Orientation and Membrane Partition Free Energy of PeT-Based Voltage-Sensitive Dyes from Molecular Simulations.

Voltage measurement via small-molecule fluorescent indicators is a valuable approach in deciphering complex dynamics in electrically excitable cells. However, our understanding of various physicochemical properties governing the performance of fluorescent voltage sensors based on the photoinduced electron transfer (PeT) mechanism remains incomplete. Here, through extensive molecular dynamics and free energy calculations, we systematically examine the orientation and membrane partition of three PeT-based voltage-sensing VoltageFluor (VF) dyes in different lipid environment. We show that the symmetry of the molecular scaffold and the net charge of the hydrophilic headgroup of a given VF dye dominate its orientation and membrane partition, respectively. Our work provides a mechanistic understanding of the physical properties contributing to the voltage sensitivity, signal-to-noise ratio, as well as membrane distribution of VF dyes and sheds light onto rational design principles of PeT-based fluorescent probes in general.

Molecular dynamics simulations of adsorption behavior of mixtures of surfactant and polymer at C3A/water and Ettringite/water interfaces

In cementitious system, surfactant is used as air-entraining agents, which is often incompatible with other chemical admixtures, such as superplasticizers. The adsorption of surfactants at solid–liquid interface is critical for the bubble stability. The measurement of adsorption properties of surfactant in fresh cement or concrete is very difficult. In this study, classical molecular dynamics simulations is used to analysis the influence of two polymers (call as HPMC and PCA, respectively) on the adsorption behaviors of the two anionic surfactants (call as HSO4 and HPO4, respectively) at the C3A/water and ettringite/water interfaces. At solid/water interface, both of HSO4 and HPO4 molecules form a monolayer with hydrophilic head groups adsorbed on the solid surfaces. After adding polymers, the adsorption conformations are changed to spherical micelle or multilayer. The electrostatic interaction between oxygen atoms of hydrophilic head groups and calcium atoms of solid surface plays a decisive role. Therefore, the value of adsorption energy of HPO4 with more negatively charged hydrophilic head groups is greater than that of HSO4. The adsorption energies of the two surfactants are weakened by the two polymers, but the adsorption energy value of HPO4 is always greater. The influence mechanism of HPMC and PCA is different. The negatively charged PCA has a strongly competitive adsorption with the surfactants. The neutral HPMC has a weak adsorption capacity but it carries many hydroxyl groups to interact with the hydrophilic head groups of surfactants.

MiRNA‐21 Inhibitor‐Loaded Cationic Liposomes Development using the Quality by Design Approach

AbstractThe success of efficiently delivering genetic material to target cells depends on developing an appropriate vector. Among the variety of nanocarriers used for gene therapy, cationic liposomes are the most investigated for delivering genetic material in different forms of cancer. Cationic liposomes contain lipids with positively charged head groups that form electrostatic bonds with the negatively charged phosphate group of nucleic acids, ensuring efficient loading of the genetic material. The aim of this study was the development and in vitro characterization of liposomal formulations for genetic material delivery using the Quality by Design (QbD) approach. We have employed the Design of Experiments (DoE) methodology to study the impact of selected formulation factors on the quality of the proposed non‐viral vectors. Among the varied factors, the ones that had an impact on the responses were the total lipid concentration and cationic lipid concentration. Two liposomal formulations with adequate critical quality attributes (CQAs) were selected for miRNA‐21 inhibitor encapsulation. We continued with the in vitro evaluation on lung cancer cell line. Fluorescence microscopy helped visualize the internalization of all liposomal formulations and apoptotic effects. Finally, we evaluated the cellular viability and migration events after treatment with the cationic liposomes.

Early events during the aggregation of Aβ16-22-derived switch-peptides tracked using Protein Charge Transfer Spectra

BackgroundUnderstanding Aβ aggregation and inhibiting it at early stages is of utmost importance in treating Alzheimer's and other related amyloidogenic diseases. However, majority of the techniques to study Aβ aggregation mainly target the late stages; while those used to monitor early stages are either expensive, use extrinsic dyes, or do not provide information on molecular level interactions. Here, we investigate the early events of Aβ16-22(KLVFFAE) aggregation using Aβ16-22 derived switch-peptides (SwPs) through a novel label-free approach employing Protein Charge Transfer Spectra (ProCharTS). ResultsWhen pH is increased from 2 to 7.2, the Aβ-derived switch peptides undergo controlled self-assembly, where the initial random coil peptides convert into β-sheet. We leveraged the intrinsic absorbance/luminescence arising from ProCharTS among growing peptide oligomers to observe the aggregation kinetics in real-time. In comparison to monomer, the lysine and glutamate headgroups in the peptide oligomer are expected to come in proximity enhancing ProCharTS intensity due to photoinduced electron transfer. With a combination of Aβ-derived switch-peptides and ProCharTS, we obtained structural insights on the early stages of Aβ-derived SwP aggregation in four unique peptides. Increase in scatter corrected ProCharTS absorbance (250–500 nm) and luminescence (320–720 nm) along with decreased mean luminescence lifetime (2.3–0.8 ns) characterize the initial stages of aggregation monitored for 1–96 h depending on the peptide. We correlated the results with Circular Dichroism (CD), 8-anilino-1-naphthalenesulfonic acid (ANS) and Thioflavin T (ThT) measurements. SignificanceWe demonstrate ProCharTS as an intrinsic analytical probe with following advantages over other conventional methods to track aggregation: it is a label-free probe; it's intensity can be measured using a UV–Vis spectrophotometer; it is more sensitive in detecting the early molecular events in aggregation compared to ANS and ThT; and it can provide information on specific contacts made between charged headgroups of Lysine/Glutamate in the oligomer.

Observation of Electrostatically Driven Surface Adsorption in Mixed Surfactant Systems.

We employed heterodyne-detected vibrational sum-frequency generation (HD-VSFG) spectroscopy to obtain a molecular-level understanding of the interaction between the anionic surfactant sodium dodecyl ammonium sulfate (SDS) and the cationic surfactant dodecyltrimethylammonium bromide (DTAB). We observed that these surfactants show a strong cooperative effect on their adsorption to the water-air interface. Even at bulk concentrations 1000 times lower than the critical micelle concentrations of SDS and DTAB, a nearly complete surface surfactant layer is observed when both surfactants are present. This strong enhancement of the surface concentrations of DS- and DTA+ can be quantitatively explained from the favorable Coulomb interaction of the oppositely charged headgroups of DS- and DTA+ and the electrostatic interactions with their counterions. The HD-VSFG results are complemented by a modified Langmuir adsorption model in which we include the free energy associated with the electrostatic interactions of the surfactant ions and their counterions.