ABSTRACT
Functionalizing nanoparticles with cell-penetrating peptides is a popular choice for cellular delivery. We investigated the effects of TAT peptide concentration and arrangement in solution on functionalized nanoparticles' efficacy for membrane permeation. We found that cell internalization correlates with the positive charge distribution achieved prior to nanoparticle encountering interactions with membrane. We identified a combination of solution based properties required to maximize the internalization efficacy of TAT-functionalized nanoparticles.
Subject(s)
Gold/chemistry , Lipid Bilayers/chemistry , Nanoparticles/chemistry , Peptides/chemistry , tat Gene Products, Human Immunodeficiency Virus/chemistry , Computer Simulation , Drug Delivery Systems , HeLa Cells , Humans , Microscopy, Electron, Transmission , Molecular Dynamics Simulation , Nanotechnology/methods , Temperature , Water/chemistryABSTRACT
The role of lipid bilayer viscoelasticity and the substrate-bilayer interactions on the spreading behavior of supported phospholipid bilayer membranes is studied using fluorescence microscopy. Unlike the monotonic roughening observed on silica or in other dynamic interface growth systems, a unique rough-smooth-rough (RSR) interface transition occurred on chromium oxide with a roughness exponent of 0.45 ± 0.04. This RSR transition is attributed to the elasticity of the lipid bilayer which is initially under compression due to surface interactions, and is well approximated by adding an elastic term to the quenched noise Edwards-Wilkinson equation. A phase diagram depicting the conditions necessary to observe RSR transitions in dynamic interface systems is derived, revealing the classes of dynamically evolving systems is broader than previously thought, and the viscoelastic nature of the lipid bilayer may play a role in supported membrane behavior.
Subject(s)
Chromium Compounds/chemistry , Lipid Bilayers/chemistry , Membrane Fluidity , Models, Chemical , Models, Molecular , Computer Simulation , Elastic Modulus , Friction , Surface PropertiesABSTRACT
Engineering the surface chemistry of a material so that it can interface with cells is an extraordinarily demanding task. The surface of a cell is composed of thousands of different lipids, proteins and carbohydrates, all intricately (and dynamically) arranged in three dimensions on multiple length scales. This complexity presents both a challenge and an opportunity to chemists working on bioactive interfaces. Here we discuss how some of these challenges can be met with interdisciplinary material synthesis. We also review the most popular classes of functional molecules grafted on engineered surfaces and explore some alternatives that may offer greater flexibility and specificity. Finally, we discuss the emerging field of dynamic surfaces capable of stimulating and responding to cellular activity in real time.
Subject(s)
Cell Membrane/chemistry , Carbohydrates/chemistry , Lipids/chemistry , Proteins/chemistryABSTRACT
Monolayer-protected metal nanoparticles (MPMNs) are a newly discovered class of nanoparticles with an ordered, striped domain structure that can be readily manipulated by altering the ratio of the hydrophobic to hydrophilic ligands. This property makes them uniquely suited to systematic studies of the role of nanostructuring on biomolecule adsorption, a phenomenon of paramount importance in biomaterials design. In this work, we examine the interaction of the simple, globular protein cytochrome C (Cyt C) with MPMN surfaces using experimental protein assays and computational molecular dynamics simulations. Experimental assays revealed that adsorption of Cyt C generally increased with increasing surface polar ligand content, indicative of the dominance of hydrophilic interactions in Cyt C-MPMN binding. Protein-surface adsorption enthalpies calculated from computational simulations employing rigid-backbone coarse-grained Cyt C and MPMN models indicate a monotonic increase in adsorption enthalpy with respect to MPMN surface polarity. These results are in qualitative agreement with experimental results and suggest that Cyt C does not undergo significant structural disruption upon adsorption to MPMN surfaces. Coarse-grained and atomistic simulations furthermore elucidated the important role of lysine in facilitating Cyt C adsorption to MPMN surfaces. The amphipathic character of the lysine side chain enables it to form close contacts with both polar and nonpolar surface ligands simultaneously, rendering it especially important for interactions with surfaces composed of adjacent nanoscale chemical domains. The importance of these structural characteristics of lysine suggests that proteins may be engineered to specifically interact with nanomaterials by targeted incorporation of unnatural amino acids possessing dual affinity to differing chemical motifs.
Subject(s)
Cytochromes c/chemistry , Metal Nanoparticles/chemistry , Molecular Dynamics Simulation , Adsorption , Cytochromes c/metabolism , Enzyme Stability , Hydrogen Bonding , Hydrophobic and Hydrophilic Interactions , Lysine , Protein Conformation , Surface Properties , ThermodynamicsABSTRACT
A flexible nanoparticle-based phospholipase (PL) assay is demonstrated in which the enzymatic substrate is decoupled from the nanoparticle surface. Liposomes are loaded with a polypeptide that is designed to heteroassociate with a second polypeptide immobilized on gold nanoparticles. Release of this polypeptide from the liposomes, triggered by PL, induces a folding-dependent nanoparticle bridging aggregation. The colorimetric response from this aggregation enables straightforward and continuous detection of PL in the picomolar range. The speed, specificity, and flexibility of this assay make it appropriate for a range of applications, from point of care diagnostics to high-throughput pharmaceutical screening.
Subject(s)
Colorimetry/methods , Liposomes/chemistry , Nanostructures/chemistry , Phospholipases/analysis , Phospholipases/chemistry , Enzyme Activation , Nanostructures/ultrastructure , Particle SizeABSTRACT
Fluid lipid bilayers were deposited on alumina substrates with the use of bubble collapse deposition (BCD). Previous studies using vesicle rupture have required the use of charged lipids or surface functionalization to induce bilayer formation on alumina, but these modifications are not necessary with BCD. Photobleaching experiments reveal that the diffusion coefficient of POPC on alumina is 0.6 microm (2)/s, which is much lower than the 1.4-2.0 microm (2)/s reported on silica. Systematically accounting for roughness, immobile regions and membrane viscosity shows that pinning sites account for about half of this drop in diffusivity. The remainder of the difference is attributed to a more tightly bound water state on the alumina surface, which induces a larger drag on the bilayer.
Subject(s)
Aluminum Oxide/analysis , Lipid Bilayers/analysis , Aluminum , Chemistry/methods , Diffusion , Equipment Design , Lipid Bilayers/chemistry , Lipids , Microscopy, Fluorescence/methods , Models, Statistical , Oxides/chemistry , Phosphatidylcholines/chemistry , Surface Properties , Time Factors , ViscosityABSTRACT
We report a new method for forming patterned lipid bilayers on solid substrates. In bubble collapse deposition (BCD), an air bubble is first "inked" with a monolayer of phospholipid molecules and then touched to the surface of a thermally oxidized silicon wafer and the air is slowly withdrawn. As the bubble shrinks, the lipid monolayer pressure increases. Once the monolayer exceeds the collapse pressure, it folds back on itself, depositing a stable lipid bilayer on the surface. These bilayer disks have lateral diffusion coefficients consistent with high quality supported bilayers. By sequentially depositing bilayers in overlapping areas, fluid connections between bilayers of different compositions are formed. Performing vesicle rupture on the open substrate surrounding this bilayer patch results in a fluid but spatially isolated bilayer. Very little intermixing was observed between the vesicle rupture and bubble-deposited bilayers.
