Using a surface forces apparatus and an atomic force microscope, we characterized the adhesive properties of adsorbed layers of two recombinant variants of Perna viridis foot protein 5 (PVFP-5), the main surface-binding protein in the adhesive plaque of the Asian green mussel. In one variant, all tyrosine residues were modified into 3,4-dihydroxy-Lphenylalanine (DOPA) during expression using a residuespecific incorporation strategy. DOPA is a key molecular moiety underlying underwater mussel adhesion. In the other variant, all tyrosine residues were preserved. The layer was adsorbed on a mica substrate and pressed against an uncoated surface. While DOPA produced a stronger adhesion than tyrosine in contact with the nanoscopic Si 3 N 4 probe of the atomic force microscope, the two variants produced comparable adhesion on the curved macroscopic mica surfaces of the surface forces apparatus. These findings show that the presence of DOPA is not a sufficient condition to generate strong underwater adhesion. Surface chemistry and contact geometry affect the strength and abundance of protein−surface bonds created during adsorption and surface contact. Importantly, the adsorbed protein layer has a random and dynamic polymer-network structure that should be optimized to transmit the tensile stress generated during surface separation to DOPA surface bonds rather than other weaker bonds.
Multiple beam interferometry (MBI) evolved as a powerful tool for the simultaneous evaluation of thin film thicknesses and refractive indices in Surface Forces Apparatus (SFA) measurements. However, analysis has relied on simplifications for providing fast or simplified analysis of recorded interference spectra. Here, we describe the implementation of new optics and a generalized fitting approach to 4 × 4 transfer matrix method simulations for the SFA. Layers are described by dispersive complex refractive indices, thicknesses, and Euler angles that can be fitted, providing modeling for birefringent or colored layers. Normalization of data by incident light intensities is essential for the implementation of a fitting approach. Therefore, a modular optical system is described that can be retrofit to any existing SFA setup. Real-time normalization of spectra by white light is realized, alignment procedures are considerably simplified, and direct switching between transmission and reflection modes is possible. A numerical approach is introduced for constructing transfer matrices for birefringent materials. Full fitting of data to the simulation is implemented for arbitrary multilayered stacks used in SFA. This enables self-consistent fitting of mirror thicknesses, birefringence, and relative rotation of anisotropic layers (e.g., mica), evaluation of reflection and transmission mode spectra, and simultaneous fitting of thicknesses and refractive indices of media confined between two surfaces. In addition, a fast full spectral fitting method is implemented for providing a possible real-time analysis with up to 30 fps. We measure and analyze refractive indices of confined cyclohexane, the thickness of lipid bilayers, the thickness of metal layers, the relative rotation of birefringent materials, contact widths, as well as simultaneous fitting of both reflection and transmission mode spectra of typical interferometers. Our analyses suggest a number of best practices for conducting SFA and open MBI in an SFA for increasingly complex systems, including metamaterials, multilayered anisotropic layers, and chiral layers.
Polymer brushes are widely used to alter the properties of interfaces. In particular, poly(ethylene glycol) (PEG) and similar polymers can make surfaces inert toward biomolecular adsorption. Neutral hydrophilic brushes are normally considered to have static properties at a given temperature. As an example, PEG is not responsive to pH or ionic strength. Here we show that, by simply introducing a polymeric acid such as poly(methacrylic acid) (PMAA), the highly hydrated brush barrier can change its properties entirely. This is caused by multivalent hydrogen bonds in an extremely pH-sensitive process. Remarkably, it is sufficient to reduce the pH to 5 for complexation to occur at the interface, which is two units higher than in the corresponding bulk systems. Below this critical pH, PMAA starts to bind to PEG in large amounts (comparable to the PEG amount), causing the brush to gradually compact and dehydrate. The brush also undergoes major rheology changes, from viscoelastic to rigid. Furthermore, the protein repelling ability of PEG is lost after reaching a threshold in the amount of PMAA bound. The changes in brush properties are tunable and become more pronounced when more PMAA is bound. The initial brush state is fully recovered when releasing PMAA by returning to physiological pH. Our findings are relevant for many applications involving functional interfaces, such as capture–release of biomolecules.
Understanding interaction force versus distance profiles of supported lipid bilayers (SLBs) is relevant to a number of areas which rely on these model systems, including e.g. characterization of ligand/receptor interactions or bacterial adhesion. Here, the stability of 4 different SLB architectures was compared using the surface forces apparatus (SFA) and atomic force microscopy (AFM). Specifically, the outer envelope of the bilayer systems remained constant as 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). The inner layer was varied between DPPC and 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP), both on mica and self-assembled monolayers (SAMs) of hexadecanethiol and the polymer tethered di-phytanylglycerol-tetraethyleneglycollipoid acid (DPhyTL) on smooth gold surfaces. In that same order these gave an increasing strength of interaction between the inner layer and the supporting substrate and hence improved stability under highly adhesive conditions. Detachment profiles from highly charged and highly adhesive contacts were characterized and approach characteristics were fitted to DLVO models. We find increasing stability under highly adhesive loads, approaching the hydrophobic limit of the adhesive energy between the inner and outer layers for the SAM based systems. For all four SLBs we further compare AFM surface topographies, which strongly depend on preparation conditions, and the DLVO fitting of the SFA approach curves find a strong charge regulation behaviour during interaction, dependent on the particular model system. In addition, we find undulation characteristics during approach and separation. The increased stability of the complex architectures on a gold support make these model systems an ideal starting point for studying more complex strongly adhesive/interacting systems, including for example ligand/receptor interactions, bio-sensing interactions or cell/surface interactions.
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