Reid C. Van Lehn

Effect of Particle Diameter and Surface Composition on the Spontaneous Fusion of Monolayer-Protected Gold Nanoparticles with Lipid Bilayers

Anionic, monolayer-protected gold nanoparticles (AuNPs) have been shown to nondisruptively penetrate cellular membranes. Here, we show that a critical first step in the penetration process is potentially the fusion of such AuNPs with lipid bilayers. Free energy calculations, experiments on unilamellar and multilamellar vesicles, and cell studies all support this hypothesis. Furthermore, we show that fusion is only favorable for AuNPs with core diameters below a critical size that depends on the monolayer composition.

Lipid tail protrusions mediate the insertion of nanoparticles into model cell membranes

Recent work has demonstrated that charged gold nanoparticles (AuNPs) protected by an amphiphilic organic monolayer can spontaneously insert into the core of lipid bilayers to minimize the exposure of hydrophobic surface area to water. However, the kinetic pathway to reach the thermodynamically stable transmembrane configuration is unknown. Here, we use unbiased atomistic simulations to show the pathway by which AuNPs spontaneously insert into bilayers and confirm the results experimentally on supported lipid bilayers. The critical step during this process is hydrophobic-hydrophobic contact between the core of the bilayer and the monolayer of the AuNP that requires the stochastic protrusion of an aliphatic lipid tail into solution. This last phenomenon is enhanced in the presence of high bilayer curvature and closely resembles the putative pre-stalk transition state for vesicle fusion. To the best of our knowledge, this work provides the first demonstration of vesicle fusion-like behaviour in an amphiphilic nanoparticle system.

Penetration of lipid bilayers by nanoparticles with environmentally-responsive surfaces: simulations and theory

Understanding the interactions between nanoparticles (NPs) and lipid bilayers is critical for the design of drug delivery carriers, biosensors, and biocompatible materials. In particular, it is desirable to understand how to effectively translocate synthetic molecules through the cellular membrane, which acts as a selective barrier to regulate transport into the cell. In this work, we use simulations and theory to explore the role that surface reconstruction may play in non-specific interactions between NPs and lipid bilayers. We show that NPs with a mixed hydrophobic/hydrophilic surface functionalization capable of rearranging their surfaces to maximize hydrophobic matching with the bilayer core are able to spontaneously establish a thermodynamically-favored position at the bilayer midplane. Furthermore, this penetration behavior is most favorable thermodynamically when the surface of the NP is near an order-disorder transition. Our analysis provides design criteria for future synthetic NPs, with the goal of designing particles that can maintain a stable transmembrane orientation.

Free energy change for insertion of charged, monolayer-protected nanoparticles into lipid bilayers

Charged, monolayer-protected gold nanoparticles (AuNPs) with core diameters smaller than 10 nm have recently emerged as a prominent class of nanomaterial for use in targeted drug delivery and biosensing. In particular, recent experimental studies showed that AuNPs protected by a binary mixture of purely hydrophobic and anionic, end-functionalized alkanethiol ligands were able to spontaneously penetrate through cell membranes via a non-endocytic, non-disruptive mechanism. The critical step in the penetration process is a fusion step during which the AuNPs insert into the hydrophobic core of the bilayer. This fusion step is driven by hydrophobic forces as inserted AuNPs minimize their exposed hydrophobic surface area and thereby lower their free energy compared to particles in the bulk. Here, we explore the effect of the large parameter space of composition, size, ligand length, morphology, and hydrophobicity strength on the change in the free energy upon insertion. Using a newly developed implicit bilayer, implicit solvent simulation model, our work shows that there is a size cutoff for insertion that has a strong dependence on surface composition and ligand chemistry. Our results agree well with previous experimental findings for a particular value of the hydrophobicity strength. This work provides physical insight that may be used to both understand the insertion of AuNPs into bilayers and guide the design of monolayers to either encourage or inhibit insertion.

Ligand-Mediated Short-Range Attraction Drives Aggregation of Charged Monolayer-Protected Gold Nanoparticles

Monolayer-protected gold nanoparticles (AuNPs) are a promising new class of nanomaterials with applications in drug delivery, self-assembly, and biosensing. The versatility of the AuNP platform is conferred by the properties of the protecting monolayer which can be engineered to tune the surface functionality of the nanoparticles. However, many applications are hampered by AuNP aggregation, which can inhibit functionality or induce particles to precipitate out of solution, even for water-soluble AuNPs. It is critical to understand the mechanisms of aggregation in order to optimally engineer protecting monolayers that both inhibit aggregation and maintain functionality. In this work, we use implicit solvent simulations to calculate the free energy change associated with the aggregation of two small, charged, alkanethiol monolayer-protected AuNPs under typical biological conditions. We show that aggregation is driven by the hydrophobic effect related to the amphiphilic nature of the alkanethiol ligands. The critical factor that enables aggregation is the deformation of ligands in the monolayer to shield hydrophobic surface area from water upon close association of the two particles. Our results further show that ligand deformation, and thus aggregation, is highly dependent on the size of the AuNPs, choice of ligands, and environmental conditions. This work provides insight into the key role that ligand-ligand interactions play in stabilizing AuNP aggregates and suggests guidelines for the design of protecting monolayers that inhibit aggregation under typical biological conditions.

Membrane-Embedded Nanoparticles Induce Lipid Rearrangements Similar to Those Exhibited by Biological Membrane Proteins

Amphiphilic monolayer-protected gold nanoparticles (NPs) have recently been shown to spontaneously fuse with lipid bilayers under typical physiological conditions. The final configuration of these NPs after fusion is proposed to be a bilayer-spanning configuration resembling transmembrane proteins. In this work, we use atomistic molecular dynamics simulations to explore the rearrangement of the surrounding lipid bilayer after NP insertion as a function of particle size and monolayer composition. All NPs studied induce local bilayer thinning and a commensurate decrease in local lipid tail order. Bilayer thickness changes of similar magnitude have been shown to drive protein aggregation, implying that NPs may also experience a membrane-mediated attraction. Unlike most membrane proteins, the exposed surface of the NP has a high charge density that causes electrostatic interactions to condense and reorient nearby lipid head groups. The decrease in tail order also leads to an increased likelihood of lipid tails spontaneously protruding toward solvent, a behavior related to the kinetic pathway for both NP insertion and vesicle-vesicle fusion. Finally, our results show that NPs can even extract lipids from the surrounding bilayer to preferentially intercalate within the exposed monolayer. These drastic lipid rearrangements are similar to the lipid mixing encouraged by fusion peptides, potentially allowing these NPs to be tuned to perform a similar biological function. This work complements previous studies on the NP-bilayer fusion mechanism by detailing the response of the bilayer to an embedded NP and suggests guidelines for the design of nanoparticles that induce controllable lipid rearrangements.

Communication: Lateral phase separation of mixed polymer brushes physisorbed on planar substrates

Here, we present a new method to model lateral phase separation in mixed polymer brushes physisorbed to a planar surface with mobile grafting points. The model is based on a local mean field theory that combines a Flory-Huggins approximation for interaction enthalpies with an Alexander-de Gennes brush entropy contribution. Using Monte Carlo sampling, the application of these two interactions to a lattice model yields a range of phase behavior consistent with previous theoretical and experimental work. This model will be useful for predicting mixed polymer brush morphologies on planar surfaces and in principle can be extended to other geometries (e.g., spheres) and polymer systems.

Universal kinetic solvent effects in acid-catalyzed reactions of biomass-derived oxygenates

The rates of Bronsted-acid-catalyzed reactions of ethyl tert-butyl ether, tert-butanol, levoglucosan, 1,2-propanediol, fructose, cellobiose, and xylitol were measured in solvent mixtures of water with three polar aprotic cosolvents: γ-valerolactone; 1,4-dioxane; and tetrahydrofuran. As the water content of the solvent environment decreases, reactants with more hydroxyl groups have higher catalytic turnover rates for both hydrolysis and dehydration reactions. We present classical molecular dynamics simulations to explain these solvent effects in terms of three simulation-derived observables: (1) the extent of water enrichment in the local solvent domain of the reactant; (2) the average hydrogen bonding lifetime between water molecules and the reactant; and (3) the fraction of the reactant accessible surface area occupied by hydroxyl groups, all as a function of solvent composition. We develop a model, constituted by linear combinations of these three observables, that predicts experimentally determined rate constants as a function of solvent composition for the entire set of acid-catalyzed reactions.

Solvent-exposed lipid tail protrusions depend on lipid membrane composition and curvature

The stochastic protrusion of hydrophobic lipid tails into solution, a subclass of hydrophobic membrane defects, has recently been shown to be a critical step in a number of biological processes like membrane fusion. Understanding the factors that govern the appearance of lipid tail protrusions is critical for identifying membrane features that affect the rate of fusion or other processes that depend on contact with solvent-exposed lipid tails. In this work, we utilize atomistic molecular dynamics simulations to characterize the likelihood of tail protrusions in phosphotidylcholine lipid bilayers of varying composition, curvature, and hydration. We distinguish two protrusion modes corresponding to atoms near the end of the lipid tail or near the glycerol group. Through potential of mean force calculations, we demonstrate that the thermodynamic cost for inducing a protrusion depends on tail saturation but is insensitive to other bilayer structural properties or hydration above a threshold value. Similarly, highly curved vesicles or micelles increase both the overall frequency of lipid tail protrusions as well as the preference for splay protrusions, both of which play an important role in driving membrane fusion. In multi-component bilayers, however, the incidence of protrusion events does not clearly depend on the mismatch between tail length or tail saturation of the constituent lipids. Together, these results provide significant physical insight into how system components might affect the appearance of protrusions in biological membranes, and help explain the roles of composition or curvature-modifying proteins in membrane fusion.

Structurally detailed coarse-grained model for Sec-facilitated co-translational protein translocation and membrane integration

We present a coarse-grained simulation model that is capable of simulating the minute-timescale dynamics of protein translocation and membrane integration via the Sec translocon, while retaining sufficient chemical and structural detail to capture many of the sequence-specific interactions that drive these processes. The model includes accurate geometric representations of the ribosome and Sec translocon, obtained directly from experimental structures, and interactions parameterized from nearly 200 μs of residue-based coarse-grained molecular dynamics simulations. A protocol for mapping amino-acid sequences to coarse-grained beads enables the direct simulation of trajectories for the co-translational insertion of arbitrary polypeptide sequences into the Sec translocon. The model reproduces experimentally observed features of membrane protein integration, including the efficiency with which polypeptide domains integrate into the membrane, the variation in integration efficiency upon single amino-acid mutations, and the orientation of transmembrane domains. The central advantage of the model is that it connects sequence-level protein features to biological observables and timescales, enabling direct simulation for the mechanistic analysis of co-translational integration and for the engineering of membrane proteins with enhanced membrane integration efficiency.