Hiroyasu Mizuno

Membrane selectivity by W-tagging of antimicrobial peptides.

A pronounced membrane selectivity is demonstrated for short, hydrophilic, and highly charged antimicrobial peptides, end-tagged with aromatic amino acid stretches. The mechanisms underlying this were investigated by a method combination of fluorescence and CD spectroscopy, ellipsometry, and Langmuir balance measurements, as well as with functional assays on cell toxicity and antimicrobial effects. End-tagging with oligotryptophan promotes peptide-induced lysis of phospholipid liposomes, as well as membrane rupture and killing of bacteria and fungi. This antimicrobial potency is accompanied by limited toxicity for human epithelial cells and low hemolysis. The functional selectivity displayed correlates to a pronounced selectivity of such peptides for anionic lipid membranes, combined with a markedly reduced membrane activity in the presence of cholesterol. As exemplified for GRR10W4N (GRRPRPRPRPWWWW-NH(2)), potent liposome rupture occurs for anionic lipid systems (dioleoylphosphatidylethanolamine (DOPE)/dioleoylphosphatidylglycerol (DOPG) and Escherichia coli lipid extract) while that of zwitterionic dioleoylphosphatidylcholine (DOPC)/cholesterol is largely absent under the conditions investigated. This pronounced membrane selectivity is due to both a lower peptide binding to the zwitterionic membranes (z≈-8-10mV) than to the anionic ones (z≈-35-40mV), and a lower degree of membrane incorporation in the zwitterionic membranes, particularly in the presence of cholesterol. Replacing cholesterol with ergosterol, thus mimicking fungal membranes, results in an increased sensitivity for peptide-induced lysis, in analogy to the antifungal properties of such peptides. Finally, the generality of the high membrane selectivity for other peptides of this type is demonstrated.

Embedded proteins and sacrificial bonds provide the strong adhesive properties of gastroliths

The adhesive properties of gastroliths from a freshwater crayfish (Cherax quadricarinatus) were quantified by colloidal probe atomic force microscopy (AFM) between heavily demineralized gastrolith microparticles and gastrolith substrates of different composition. Combined AFM and transmission electron microscopy studies demonstrated that the sequential detachment and large adhesion energies that characterise the adhesive behaviour of a native gastrolith substrate are dominated by sacrificial bonds between chitin fibres and between chitin fibres and CaCO(3). The sacrificial bonds were shown to be strongly related to the gastrolith proteins and when the majority of these proteins were removed by ethylenediaminetetraacetic acid (EDTA), the sequential detachment disappeared and the adhesive energy was reduced by more than two orders of magnitude.

Friction Measurement Between Polyester Fibres Using the Fibre Probe SPM

An SPM has been used to measure frictional interactions between two crossed fibres for the first time. The preparation of the surfaces is briefly described, but the crucial element is that the fibr ...

New Insight on the Friction of Natural Fibers. Effect of Sliding Angle and Anisotropic Surface Topography

The friction anisotropy of human hair has been investigated as a function of angle using AFM fiber probe measurements to evaluate the role of cuticle alignment. It is found that friction hysteresis, the difference in friction coefficients between sliding with or against the cuticle direction, is essentially nonexistent for native human hair. For damaged human hair, however, a clear friction hysteresis is observed, which appears to be a periodic function of the angle between the fibers. The implication is that antiparallel sliding is not in itself sufficient for friction isotropy but that lifting of the cuticle edges is required. A methodology to perform friction analysis independently for trace and retrace was therefore developed, which is applicable to any type of AFM lateral force measurement. It explicitly accounts for roll, noncircular cross section, and off-axis alignment as well as baseline drift, which allows real anisotropy in the friction coefficient to be deconvoluted from these artifacts.