Biomedical subjects

Per Lyngs Hansen

Publications and source records attributed to Per Lyngs Hansen.

5 recordsLinked to original sources

PubMed2006

Dynamic force spectroscopy on soft molecular systems: improved analysis of unbinding spectra with varying linker compliance.

Dynamic force spectroscopy makes it possible to measure the breaking of single molecular bonds or the unfolding of single proteins subjected to a time-dependent pulling force. The force needed to break a single bond or to unfold a domain in a protein depends critically on the time dependence of the applied force. In this way the elastic response couples to the unbinding force. We have performed an experimental and theoretical examination of this coupling by studying the well-known biotin-streptavidin bond in systems incorporating two common types of linkers. In the first case biotin is linked by bovine serum albumin (BSA) and it is observed that this linker has a linear elastic response. More surprisingly we find that its force constant varies significantly between repeated force curves. It is demonstrated that by sorting the force curves according to the force constant of the linker we can improve the data analysis and obtain a better agreement between experimental data and theory. In the second case biotin is linked by poly(ethylene glycol) (PEG), which has a soft nonlinear elastic response. A numerical calculation of the unbinding statistics for the polymer system agrees quantitatively with experiments. It demonstrates a clear decrease in unbinding forces resulting from the polymer linker.

Animals↗

PubMed2006

Hydrolysis of fluid supported membrane islands by phospholipase A(2): Time-lapse imaging and kinetic analysis.

The activity of phospholipase A(2) (PLA(2)) which catalyzes the hydrolysis of phospholipids into free fatty acids and lysolipids, depends on the structure and thermodynamic state of the membrane. To further understand how the substrate conformation correlates with enzyme activity, model systems that are based on time-resolved membrane microscopy are needed. We demonstrate a methodology for preparing and investigating the dynamics of fluid supported phospholipid membranes hydrolyzed by snake venom PLA(2). The method uses quantitative analysis of time-lapse fluorescence images recording the evolution of fluid bilayer islands during hydrolysis. In order to minimize interactions with the support surface, we use double bilayer islands situated on top of a complete primary supported membrane prepared by hydration of spincoated lipid films. Our minimal kinetic analysis describes adsorption of enzyme to the membrane in terms of the Langmuir isotherm as well as enzyme kinetics. We use two related models assuming hydrolysis to occur either at the perimeter or at the surface of the membrane island. We find that the adsorption constant is similar for the two cases, while the estimated turnover rate is markedly different. The PLA(2) concentration series is measured in the absence and presence of beta-cyclodextrin which forms water soluble complexes with the reaction products. The results demonstrate the versatility of double bilayer islands as a membrane model system and introduces a new method for quantifying the kinetics of lipase activity on membranes by directly monitoring the evolution in substrate morphology.

Adsorption↗

PubMed2005

DNA overstretching transition: ionic strength effects.

As double-stranded DNA is stretched to its B-form contour length, models of polymer elasticity can describe the dramatic increase in measured force. When the molecule is stretched beyond the contour length, it further shows a highly cooperative overstretching transition. We have developed a theoretical description for this transition by coupling the two-state model and the elasticity theory proposed earlier by others. Furthermore, we have extended this model to account for monovalent salt effects on elastic moduli during the transition. We find that this theoretical description is in very good agreement with recent measurements for the salt dependence of the overstretching transition, allowing us to gain insight into the mechanisms that govern the transition. In double-stranded DNA, the effective length per unit charge varies with salt in agreement with the Manning and Poisson-Boltzmann models for thin polyelectrolyte rods, whereas the other model parameters describing structural features have barely any salt dependence. The results thus suggest that the electrostatic component of force-induced overstretching is mediated mesoscopically via elasticity.

Computer Simulation↗

PubMed2004

Nanobubbles give evidence of incomplete wetting at a hydrophobic interface.

The appearance of a hydrophobic surface, namely a crystalline (111) Si wafer coated with a thick soft polystyrene film, and the morphological changes along this interface depending on the polarity of an adjoining liquid phase were studied with magnetic tapping mode atomic force microscopy. Interfacially associated nanobubbles of decreasing size and number are observed as the hydrophobicity of the subphase increases. The disturbance of the water structure in the contact region induces the formation of nanobubbles. The topology of the interface is visualized, starting with the dry polymer under normal atmosphere conditions and observing the changes as air is replaced by a series of liquids. With water, the surface coverage of the substrate with bubbles is almost a close-packed configuration. The bubble shape is well approximated by spherical caps of a rather low aspect ratio. The Gaussian size distributions of bubble shape parameters are discussed. The contact angle of the nanobubbles is substantially smaller than the corresponding number measured for a macroscopic droplet. This apparent discrepancy might be resolved if the nanobubbles were assumed to exist along the interface as a connecting sublayer between a depleted water film at the hydrophobic polymer surface and an adsorbed macrodroplet.

Journal Article↗

PubMed2003

Osmotic properties of poly(ethylene glycols): quantitative features of brush and bulk scaling laws.

From glycosylated cell surfaces to sterically stabilized liposomes, polymers attached to membranes attract biological and therapeutic interest. Can the scaling laws of polymer "brushes" describe the physical properties of these coats? We delineate conditions where the Alexander-de Gennes theory of polymer brushes successfully fits the intermembrane distance versus applied osmotic stress data of Kenworthy et al. for poly(ethylene glycol)-grafted multilamellar liposomes. We establish that the polymer density and size in the brush must be high enough that, in a bulk solution of equivalent monomer density, the polymer osmotic pressure is independent of polymer molecular weight (the des Cloizeaux semidilute regime of bulk polymer solutions). The condition that attached polymers behave as semidilute bulk solutions offers a rigorous criterion for brush scaling-law behavior. There is a deep connection between the behaviors of semidilute polymer solutions in bulk and polymers grafted to a surface at a density such that neighbors pack to form a uniform brush. In this regime, two-parameter unconstrained fits of the Alexander-de Gennes brush scaling laws to the Kenworthy et al. data yield effective monomer lengths of 3.3-3.6 A, which agree with structural predictions. The fitted distances between grafting sites are larger than expected from the nominal mole fraction of poly(ethylene glycol)-lipids; the chains apparently saturate the surface. Osmotic stress measurements can be used to estimate the actual densities of membrane-grafted polymers.

Computer Simulation↗