α-Synuclein Senses Lipid Packing Defects and Induces Lateral Expansion of Lipids Leading to Membrane Remodeling

There is increasing evidence for the involvement of lipid membranes in both the functional and pathological properties of α-synuclein (α-Syn). Despite many investigations to characterize the binding of α-Syn to membranes, there is still a lack of understanding of the binding mode linking the properties of lipid membranes to α-Syn insertion into these dynamic structures. Using a combination of an optical biosensing technique and in situ atomic force microscopy, we show that the binding strength of α-Syn is related to the specificity of the lipid environment (the lipid chemistry and steric properties within a bilayer structure) and to the ability of the membranes to accommodate and remodel upon the interaction of α-Syn with lipid membranes. We show that this interaction results in the insertion of α-Syn into the region of the headgroups, inducing a lateral expansion of lipid molecules that can progress to further bilayer remodeling, such as membrane thinning and expansion of lipids out of the membrane plane. We provide new insights into the affinity of α-Syn for lipid packing defects found in vesicles of high curvature and in planar membranes with cone-shaped lipids and suggest a comprehensive model of the interaction between α-Syn and lipid bilayers. The ability of α-Syn to sense lipid packing defects and to remodel membrane structure supports its proposed role in vesicle trafficking.     

FlashPack: Fast and Simple Preparation of Ultrahigh-performance Capillary Columns for LC-MS*

1. Download : Download high-res image (99KB)
2. Download : Download full-size image

Highlights

• •Fast and simple capillary column packing protocol.
• •Low-pressure packing at <100 bars from ultrahigh sorbent suspension concentration.
• •Sorbent particle aggregation leading to blocking of the column entrance is avoided.
• •Effective for long capillary UHPLC column packing with a wide range of sorbents.

     

Starch analysis using hydrodynamic chromatography with a mixed-bed particle column

Columns packed with commercial glass beads 5 and 19 μm average size and a mixture of both (0.7 volume fraction of large particles) were used to analyse starch composition by hydrodynamic chromatography (HDC), applying water as mobile phase. To obviate retrogradation, experiments were carried out at column temperatures of 15 and 3 °C and several types of starch were assayed. In what concerns amylopectin and amylose separation, a better resolution and a lower pressure drop were obtained for the mixed binary packing when compared with the packing containing uniform 5 μm glass beads. A more efficient cooling of the mobile phase was also obtained with the mixed packing, which was determinant for improving resolution. For the Hylon VII starch the relative retention times (RRT) were 0.777 and 0.964 for amylopectin and amylose, respectively, while for the Tapioca starch the obtained RRTs were 0.799 and 0.923. Application of unbound glass beads as column packing not only might reduce equipment and running costs in preparative scale separations, but also proved to be useful as a fast and reliable method to monitor the amylose and amylopectin content of starch samples of different sources.     

Human endothelial cells contain one type of plasminogen activator

RNA-binding protein kinase from amphibian oocytes modifies serine and threonine residues in the molecules of substrates and utilizes both ATP and GTP. Low concentrations of heparin inhibit protein kinase. The foregoing suggests that this enzyme is casein kinase II. It is shown that RNA-binding proteins lack active forms of phosphatases and proteases which may affect the results of phosphorylation of both endogenous and exogenous substrates.     

Parameters modulating the maximum insertion pressure of proteins and peptides in lipid monolayers

The lipid monolayer model membrane is useful for studying the parameters responsible for protein and peptide membrane binding. Different approaches have been used to determine the extent of protein and peptide binding to lipid monolayers. This review focuses on the use of the “maximum insertion pressure” (MIP) to estimate the extent of protein and peptide penetration in lipid monolayers. The MIP data obtained with different proteins and peptides have been reviewed and discussed which allowed to draw conclusions on the parameters modulating the monolayer binding of proteins and peptides. In particular, secondary structure components such as amphipathic α-helices of proteins and peptides as well as electrostatic interactions play important roles in monolayer binding. The MIPs have been compared to the estimated lateral pressure of biomembranes which allowed to evaluate the possible association between proteins or peptides with natural membranes. For example, the MIP of a membrane-anchored protein with a glycosylphosphatidylinositol (GPI) was found to be far below the estimated lateral pressure of biomembranes. This allowed us to conclude that this protein is probably unable to penetrate the membrane and should thus be hanged at the membrane surface by use of its GPI lipid anchor. Moreover, the values of MIP obtained with myristoylated and non-myristoylated forms of calcineurin suggest that the myristoyl group does not contribute to monolayer binding. However, the acylation of a peptide resulted in a large increase of MIP. Finally, the physical state of lipid monolayers can have a strong effect on the values of MIP such that it is preferable to perform measurements with lipids showing a single physical state. Altogether the data show that the measurement of the maximum insertion pressure provides very useful information on the membrane binding properties of proteins and peptides although uncertainties must be provided to make sure the observed differences are significant.     

Finite element modeling of embolic coil deployment: Multifactor characterization of treatment effects on cerebral aneurysm hemodynamics

Endovascular coiling is the most common treatment for cerebral aneurysms. During the treatment, a sequence of embolic coils with different stiffness, shapes, sizes, and lengths is deployed to fill the aneurysmal sac. Although coil packing density has been clinically correlated with treatment success, many studies have also reported success at low packing densities, as well as recurrence at high packing densities. Such reports indicate that other factors may influence treatment success. In this study, we used a novel finite element approach and computational fluid dynamics (CFD) to investigate the effects of packing density, coil shape, aneurysmal neck size, and parent vessel flow rate on aneurysmal hemodynamics. The study examines a testbed of 80 unique CFD simulations of post-treatment flows in idealized basilar tip aneurysm models. Simulated coil deployments were validated against in vitro and in vivo deployments. Among the investigated factors, packing density had the largest effect on intra-aneurysmal velocities. However, multifactor analysis of variance showed that coil shape can also have considerable effects, depending on packing density and neck size. Further, linear regression analysis showed an inverse relationship between mean void diameter in the aneurysm and mean intra-aneurysmal velocities, which underscores the importance of coil distribution and thus coil shape. Our study suggests that while packing density plays a key role in determining post-treatment hemodynamics, other factors such as coil shape, aneurysmal geometry, and parent vessel flow may also be very important.     

Manipulation and Molecular Resolution of a Phosphatidylcholine-Supported Planar Bilayer by Atomic Force Microscopy

The morphology of supported planar bilayers has been investigated below phase transition temperature by atomic force microscopy in contact and tapping mode. The bilayers were formed by the vesicle-spreading technique. In contact mode at low scanning forces of about 1 nN true molecular resolution could be achieved for supported phosphatidylcholine bilayers. The resolution was confirmed by experiments that captured the location, average area of individual lipid headgroups and the manipulation of the bilayer surface. Repeated scanning in contact mode shifted the random topology of the surface consecutively to a striped pattern. Height profiles of defect-containing bilayers were analyzed. The shape of the defects became smooth by repeated scanning. The height profiles allowed the estimation of the indentation of the tip into the surface-adsorbed membrane. In tapping mode a disordered pattern of headgroups became visible. Our morphological data at molecular resolution suggest that the native arrangement of the choline headgroups is disordered, free of large packing defects and becomes ordered in Schallamach waves by scanning in contact mode. Received: 12 March 1997/Revised: 3 October 1997     

Morphological assembly mechanisms in Neotropical bat assemblages and ensembles within a landscape

Empirical studies on bat assemblages have shown that richness is not appreciably influenced by local processes such as ecological interactions. However, most of these studies have been done in large areas that include high heterogeneity, and they analyse all bat species within such areas, and thus they may be not reflecting local but supra-community conditions. We followed an ecomorphological approach to assess how bat assemblages of species from the families Phyllostomidae and Mormoopidae, and ensembles of frugivorous bats, are assembled in local habitats within a single landscape. We measured the volume of the space defined by wing morphology and quantified the average distance between species within such a volume. Then, we related these measures to local richness. Such relationships were contrasted against relationships with random assemblages to test for statistical differences. At the ensemble level of organization, we found that the frugivorous bat morphological assembly mechanism is different from random patterns, and it corresponds to the volume-increasing model. On the other hand, bat assembly mechanisms may be ubiquitous at the assemblage level, because groups of species coexisting in a local habitat and delimited only by phylogeny include more than one ecological group with no potential to interact. Assembling processes are crucial to an understanding of species diversity in local communities, and ecomorphological analyses are very promising tools that may help in their study.     

Heterogeneity in iota-carrageenan molecular structure: insights for polymorph II-->III transition in the presence of calcium ions

Iota-carrageenan is used in pharmaceutical and food applications due to its ability to complex with other hydrocolloids and proteins. Six distinct cation dependent allomorphs, consistent with its versatile functionality, have so far been observed in the solid state. In this contribution, X-ray structural details of calcium iota-carrageenan (form III) are reported. The polysaccharide retains the half-staggered, parallel, 3-fold, right-handed double helix stabilized by interchain hydrogen bonds from O-2H and O-6H in the Galp units. Results show that there are four helices, rather than one in I or three in II, organized in a larger pseudo-trigonal unit cell of dimensions a=27.44, c=13.01 A, and gamma=120 degrees . The four helices have similar core structures, but their sulfate group orientations are quite different. Fifteen calcium ions and 64 water molecules hold the helices together and promote helix-helix interactions. The results portray how the helices would shuffle around in an orchestrated manner to yield calcium iota-carrageenan III from II.     

Influence of synthetic packing materials on the gas dispersion and biodegradation kinetics in fungal air biofilters

The biodegradation of toluene was studied in two lab-scale air biofilters operated in parallel, packed respectively with perlite granules (PEG) and polyurethane foam cubes (PUC) and inoculated with the same toluene-degrading fungus. Differences on the material pore size, from micrometres in PEG to millimetres in PUC, were responsible for distinct biomass growth patterns. A compact biofilm was formed around PEG, being the interstitial spaces progressively filled with biomass. Microbial growth concentrated at the core of PUC and the excess of biomass was washed-off, remaining the gas pressure drop comparatively low. Air dispersion in the bed was characterised by tracer studies and modelled as a series of completely stirred tanks (CSTR). The obtained number of CSTR (n) in the PEG packing increased from 33 to 86 along with the applied gas flow (equivalent to empty bed retention times from 48 to 12 s) and with operation time (up to 6 months). In the PUC bed, n varied between 9 and 13, indicating that a stronger and steadier gas dispersion was achieved. Michaelis–Menten half saturation constant (k _m) estimates ranged 71–113 mg m⁻³, depending on the experimental conditions, but such differences were not significant at a 95% confidence interval. The maximum volumetric elimination rate (r _m) varied from 23 to 50 g m⁻³ h⁻¹. Comparison between volumetric and biomass specific biodegradation activities indicated that toluene mass transfer was slower with PEG than with PUC as a consequence of a smaller biofilm surface and to the presence of larger zones of stagnant air.