Membrane Phase Transitions in Lipid-Wrapped Nanoparticles.

A Landau theory is constructed for the gel/fluid transition of a lipid bilayer wrapped around a spherical nanoparticle (lipid-wrapped nanoparticle, LNP). The bilayer is regarded as a regular solution of gel and fluid lipids with distinct inter- and intralayer interactions plus the interaction of the core with the inner layer. It is required that both the inner and the outer surfaces of the bilayer are perfectly covered with lipids, with the gel and fluid lipids having different areas/lipid. The equilibrium state is found by minimizing the free energy as a function of the fractions of fluid lipids in the inner and outer layers. The transition has been studied extensively for lamellar membranes in the thermodynamic limit. LNP have significant curvature and are not in the thermodynamic limit. The increase of the gel energy with curvature, identified in our previous work as its most important effect, is included. The focus of the paper is the dependence of the transition on the core radius, R, controlling curvature, and the core-lipid interaction. With decreasing R, trends found in experiment are reproduced in a model calculation: (1) decrease of the transition temperature, Tm, (2) decoupling of the transitions in the inner and outer layers, and (3) possibility of lower Tm in the inner layer. The disruption of gel packing by curvature and the interaction of the core with the inner layer are highlighted as the most important determinants of deviation from bulk behavior.

Moltemplate: A Tool for Coarse-Grained Modeling of Complex Biological Matter and Soft Condensed Matter Physics.

Coarse-grained models have long been considered indispensable tools in the investigation of biomolecular dynamics and assembly. However, the process of simulating such models is arduous because unconventional force fields and particle attributes are often needed, and some systems are not in thermal equilibrium. Although modern molecular dynamics programs are highly adaptable, software designed for preparing all-atom simulations typically makes restrictive assumptions about the nature of the particles and the forces acting on them. Consequently, the use of coarse-grained models has remained challenging. Moltemplate is a file format for storing coarse-grained molecular models and the forces that act on them, as well as a program that converts moltemplate files into input files for LAMMPS, a popular molecular dynamics engine. Moltemplate has broad scope and an emphasis on generality. It accommodates new kinds of forces as they are developed for LAMMPS, making moltemplate a popular tool with thousands of users in computational chemistry, materials science, and structural biology. To demonstrate its wide functionality, we provide examples of using moltemplate to prepare simulations of fluids using many-body forces, coarse-grained organic semiconductors, and the motor-driven supercoiling and condensation of an entire bacterial chromosome.

Simulation of fluid/gel phase equilibrium in lipid vesicles.

Simulation of single component dipalmitoylphosphatidylcholine (DPPC) coarse-grained DRY-MARTINI lipid vesicles of diameter 10 nm (1350 lipids), 20 nm (5100 lipids) and 40 nm (17 600 lipids) is performed using statistical temperature molecular dynamics (STMD), to study finite size effects upon the order-disorder gel/fluid transition. STMD obtains enhanced sampling using a generalized ensemble, obtaining a flat energy distribution between upper and lower cutoffs, with little computational cost over canonical molecular dynamics. A single STMD trajectory of moderate length is sufficient to sample 20+ transition events, without trapping in the gel phase, and obtain well averaged properties. Phase transitions are analyzed via the energy-dependence of the statistical temperature, TS(U). The transition temperature decreases with decreasing diameter, in agreement with experiment, and the transition changes from first order to borderline first-second order. The size- and layer-dependence of the structure of both stable phases, and of the pathway of the phase transition, are determined. It is argued that the finite size effects are primarily caused by the disruption of the gel packing by curvature. Inhomogeneous states with faceted gel patches connected by unusual fluid seams are observed at high curvature, with visually different structure in the inner and outer layers due to the different curvatures. Thus a simple physical picture describes phase transitions in nanoscale finite systems far from the thermodynamic limit.

Membrane-wrapped nanoparticles probe divergent roles of GM3 and phosphatidylserine in lipid-mediated viral entry pathways.

Gold nanoparticles (NPs) wrapped in a membrane can be utilized as artificial virus nanoparticles (AVNs) that combine the large nonblinking or bleaching optical cross-section of the NP core with the biological surface properties and functionalities provided by a self-assembled lipid membrane. We used these hybrid nanomaterials to test the roles of monosialodihexosylganglioside (GM3) and phosphatidylserine (PS) for a lipid-mediated targeting of virus-containing compartments (VCCs) in macrophages. GM3-presenting AVNs bind to CD169 (Siglec-1)-expressing macrophages, but inclusion of PS in the GM3-containing AVN membrane decreases binding. Molecular dynamics simulations of the AVN membrane and experimental binding studies of CD169 to GM3-presenting AVNs reveal Na+-mediated interactions between GM3 and PS as a potential cause of the antagonistic action on binding by the two negatively charged lipids. GM3-functionalized AVNs with no or low PS content localize to tetherin+, CD9+ VCC in a membrane composition-depending fashion, but increasing amounts of PS in the AVN membrane redirect the NP to lysosomal compartments. Interestingly, this compartmentalization is highly GM3 specific. Even AVNs presenting the related monosialotetrahexosylganglioside (GM1) fail to achieve an accumulation in VCC. AVN localization to VCC was observed for AVN with gold NP core but not for liposomes, suggesting that NP sequestration into VCC has additional requirements beyond ligand (GM3)-receptor (CD169) recognition that are related to the physical properties of the NP core. Our results confirm AVN as a scalable platform for elucidating the mechanisms of lipid-mediated viral entry pathways and for selective intracellular targeting.

Lipid Packing in Lipid-Wrapped Nanoparticles.

Equilibrium simulations of lipid-wrapped nanoparticles (LNP) were performed using a hybrid molecular dynamics/Monte Carlo (MD/MC) approach. The radius, R, of a spherical nanoparticle (NP) core was adjusted with MC moves while a surrounding lipid bilayer was treated with MD. A wide range of LNP sizes, with the largest R ∼ 40 nm, were studied to determine the average NP radius for a given total number of lipids, N, the number of lipids in each layer, and configurational information. A three-bead lipid model was used to allow large N. A nonequilibrium Jarzynski free energy calculation of the optimal R for a given N, was also demonstrated validating the MD/MC method. An order/disorder transition was described, unique to LNP and distinct from lamellar bilayers, that is weak and continuous with small N, but sharpens to a first order transition with N > 10000 at T ≈ 1.1, shifting to higher T with increasing N. The radius and the overlap of the inner and outer layers were used as order parameters charactering the whole system, and the density vs distance from the origin served to describe the transition in individual layers. The ordering effect of the core on the inner layer, and the disordering effect of curvature, are evident. Excellent fits for the number of lipids in the inner and outer layers vs R are presented, based on the idea that the inner layer is described as usual by an area and area/lipid but that the outer layer is slaved to the inner. The most ordered states exhibit interdigitation of the inner head groups with themselves.

Membrane Fluidity Sensing on the Single Virus Particle Level with Plasmonic Nanoparticle Transducers.

Viral membranes are nanomaterials whose fluidity depends on their composition, in particular, the cholesterol (chol) content. As differences in the membrane composition of individual virus particles can lead to different intracellular fates, biophysical tools capable of sensing the membrane fluidity on the single-virus level are required. In this manuscript, we demonstrate that fluctuations in the polarization of light scattered off gold or silver nanoparticle (NP)-labeled virus-like-particles (VLPs) encode information about the membrane fluidity of individual VLPs. We developed plasmonic polarization fluctuation tracking microscopy (PFTM) which facilitated the investigation of the effect of chol content on the membrane fluidity and its dependence on temperature, for the first time on the single-VLP level. Chol extraction studies with different methyl-ß-cyclodextrin (MßCD) concentrations yielded a gradual decrease in polarization fluctuations as a function of time. The rate of chol extraction for individual VLPs showed a broad spread, presumably due to differences in the membrane composition for the individual VLPs, and this heterogeneity increased with decreasing MßCD concentration.

Enhanced Sampling of Phase Transitions in Coarse-Grained Lipid Bilayers.

Freezing and melting of dipalmitoylphosphatidylcholine (DPPC) bilayers are simulated in both the explicit (Wet) and implicit solvent (Dry) coarse-grained MARTINI force fields with enhanced sampling, via the isobaric, molecular dynamics version of the generalized replica exchange method (gREM). Phase transitions are described with the entropic viewpoint, based upon the statistical temperature as a function of enthalpy, TS(H) = 1/(dS(H)/dH), where S is the configurational entropy. Bilayer thickness, area per lipid, and the second-rank order parameter (P2) are calculated vs temperature in the transition range. In a 32-lipid Wet MARTINI system, transitions in the lipid and water subsystems are strongly coupled, giving rise to considerable structure in TS(H) and the need to specify the state of the water when reporting a lipid transition temperature. For gel lipid + liquid water → fluid lipid + liquid water, we find 292.4 K. The small system is influenced by finite-size effects, but it is argued that the entropic approach is well suited to revealing them, which will be particularly relevant for studies of finite nanosystems where there is no thermodynamic limit. In a 390-lipid Dry MARTINI system, two-dimensional analogues of the topographies of coexisting states ("subphases") seen in pure fluids are found. They are not seen in the 32-lipid Wet or Dry system, but the Dry lipids show a new type of state with gel in one leaflet and tilted gel in the other. Dry bilayer transition temperatures are 333.3 K (390 lipids) and 338 K (32 lipids), indicating that the 32-lipid system is not too small for a qualitative study of the transition. Physical arguments are given for Dry lipid system size dependence and for the difference between Wet and Dry systems.

Hole wave functions and transport with deazaadenines replacing adenines in DNA.

Transport of a hole along the base stack of DNA is relatively facile for a series of adenines (As) paired with thymines (Ts) or for a series of guanines (Gs) paired with cytosines (Cs). However, the speed at which a hole was found to travel was much too small to make useful semiconductor-type devices. Quite recently it was found that replacing one of the electronegative nitrogens (N3 or N7) with a carbon and a hydrogen, thus turning A into deazaadenine, increased the hole speed in what was A/T by a factor 30. To study the effect of the substitution we have carried out simulations for the wave function of a hole on an A/T oligomer with As modified by replacing N3 or N7, or both, with C-H's. The simulations were carried out using QM/MM and the code CP2K. We find, for either N, or both, replaced, the wave function of the hole behaves similarly to that of a hole on A/T in being delocalized immediately after hole insertion for up to ∼20 fs, and then becoming localized on one of the modified As. The time for localization could be decreased by placing additional water within ∼1.8 Šof N3 or N7, encouraging the formation of hydrogen bonds with these nitrogens. Because of their positive charge the hydrogen bonds tend to repel holes. However, these bonds were found to decay on a femtosecond time scale, thus unlikely to affect the hole hopping, which occurs on approximately a nanosecond scale in A/T. Replacement with a C-H of one or both of the electronegative Ns, along with the structural changes that result, is expected to decrease the activation energy and thus account for the larger hole hopping rate in the deaza-modified DNA.