Autoinhibition of Endophilin in Solution via Interdomain Interactions

Endophilin is a key protein involved in clathrin-mediated endocytosis. Previous computational and experimental work suggested that the N-terminal helix is embedded into the membrane to induce curvature; however, the role of the SH3 domain remains controversial. To address this issue, we performed computer simulations of the endophilin dimer in solution to understand the interaction between the N-BAR and SH3 domains and its effect on biological function. We predict that the helix binds to the SH3 domain through hydrophobic and salt-bridge interactions. This protects the hydrophobic residues on both domains and keeps the SH3 domain near the end of the N-BAR domain, in agreement with previous experimental results. The complex has a binding strength similar to a few hydrogen bonds (13.0 ± 0.6 kcal/mol), and the SH3 domain stabilizes the structure of the N-terminal helix in solution. Electrostatic calculations show a large region of strongly positive electrostatic potential near the N-terminal that can orient the helix toward the membrane and likely embed the helix into the membrane surface. This predicted mechanism suggests that endophilin can select for both curvature and electrostatic potential when interacting with membranes, highlighting the importance of the SH3 domain in regulating the function of endophilin.     

Two conformational states of the membrane-associated Bacillus thuringiensis Cry4Ba delta-endotoxin complex revealed by electron crystallography: implications for toxin-pore formation

The insecticidal nature of Cry delta-endotoxins produced by Bacillus thuringiensis is generally believed to be caused by their ability to form lytic pores in the midgut cell membrane of susceptible insect larvae. Here we have analyzed membrane-associated structures of the 65-kDa dipteran-active Cry4Ba toxin by electron crystallography. The membrane-associated toxin complex was crystallized in the presence of DMPC via detergent dialysis. Depending upon the charge of the adsorbed surface, 2D crystals of the oligomeric toxin complex have been captured in two distinct conformations. The projection maps of those crystals have been generated at 17A resolution. Both complexes appeared to be trimeric; as in one crystal form, its projection structure revealed a symmetrical pinwheel-like shape with virtually no depression in the middle of the complex. The other form revealed a propeller-like conformation displaying an obvious hole in the center region, presumably representing the toxin-induced pore. These crystallographic data thus demonstrate for the first time that the 65-kDa activated Cry4Ba toxin in association with lipid membranes could exist in at least two different trimeric conformations, conceivably implying the closed and open states of the pore.     

Microalgal hydrogen production: prospects of an essential technology for a clean and sustainable energy economy

Modern energy production is required to undergo a dramatic transformation. It will have to replace fossil fuel use by a sustainable and clean energy economy while meeting the growing world energy needs. This review analyzes the current energy sector, available energy sources, and energy conversion technologies. Solar energy is the only energy source with the potential to fully replace fossil fuels, and hydrogen is a crucial energy carrier for ensuring energy availability across the globe. The importance of photosynthetic hydrogen production for a solar-powered hydrogen economy is highlighted and the development and potential of this technology are discussed. Much successful research for improved photosynthetic hydrogen production under laboratory conditions has been reported, and attempts are underway to develop upscale systems. We suggest that a process of integrating these achievements into one system to strive for efficient sustainable energy conversion is already justified. Pursuing this goal may lead to a mature technology for industrial deployment.     

Differentially oriented populations of actin filaments generated in lamellipodia collaborate in pushing and pausing at the cell front

Eukaryotic cells advance in phases of protrusion, pause and withdrawal. Protrusion occurs in lamellipodia, which are composed of diagonal networks of actin filaments, and withdrawal terminates with the formation of actin bundles parallel to the cell edge. Using correlated live-cell imaging and electron microscopy, we have shown that actin filaments in protruding lamellipodia subtend angles from 15-90 degrees to the front, and that transitions from protrusion to pause are associated with a proportional increase in filaments oriented more parallel to the cell edge. Microspike bundles of actin filaments also showed a wide angular distribution and correspondingly variable bilateral polymerization rates along the cell front. We propose that the angular shift of filaments in lamellipodia serves in adapting to slower protrusion rates while maintaining the filament densities required for structural support; further, we suggest that single filaments and microspike bundles contribute to the construction of the lamella behind and to the formation of the cell edge when protrusion ceases. Our findings provide an explanation for the variable turnover dynamics of actin filaments in lamellipodia observed by fluorescence speckle microscopy and are inconsistent with a current model of lamellipodia structure that features actin filaments branching at 70 degrees in a dendritic array.     

Combined MRI–PET dissects dynamic changes in plant structures and functions

Unravelling the factors determining the allocation of carbon to various plant organs is one of the great challenges of modern plant biology. Studying allocation under close to natural conditions requires non-invasive methods, which are now becoming available for measuring plants on a par with those developed for humans. By combining magnetic resonance imaging (MRI) and positron emission tomography (PET), we investigated three contrasting root/shoot systems growing in sand or soil, with respect to their structures, transport routes and the translocation dynamics of recently fixed photoassimilates labelled with the short-lived radioactive carbon isotope ¹¹C. Storage organs of sugar beet ( Beta vulgaris ) and radish plants ( Raphanus sativus ) were assessed using MRI, providing images of the internal structures of the organs with high spatial resolution, and while species-specific transport sectoralities, properties of assimilate allocation and unloading characteristics were measured using PET. Growth and carbon allocation within complex root systems were monitored in maize plants ( Zea mays ), and the results may be used to identify factors affecting root growth in natural substrates or in competition with roots of other plants. MRI–PET co-registration opens the door for non-invasive analysis of plant structures and transport processes that may change in response to genomic, developmental or environmental challenges. It is our aim to make the methods applicable for quantitative analyses of plant traits in phenotyping as well as in understanding the dynamics of key processes that are essential to plant performance.     

Spherical classification of wavelet transformed EMG intensity patterns

Electromyograms of different muscles can be submitted to a wavelet-transform and arranged in a Multi-Muscle Pattern (MMP). The MMP represents the intensity of the EMG signals of a number of muscles simultaneously in time/frequency space. As previously shown, the MMPs can be represented by points in an Euclidian vector space that was called pattern space. The variability of the MMPs is represented by the distribution of the scattered points in pattern space. The purpose of this study was to investigate the distribution of the points and use the properties of the distribution to classify MMPs. The first task was to test whether the points representing a group of MMPs were located between the inner and outer boundary of a sphere-like domain in whitened pattern space as theoretically predicted. The mean of these points and thus of the MMPs is represented by a point at the center of the sphere. The hypothesis was that the spheres representing points of the MMPs of barefoot and shod runners were sufficiently separated and distinguishable in pattern space to allow classification of the runners according to their shod condition. The results confirmed the hypothesis and revealed that the recognition rate was over 80%. One can conclude and generalize that the points representing MMPs recorded for a certain condition reside between the inner and outer boundary of the sphere. The classification based on the spherical feature represents a much better discrimination than one based on the distance from the mean.     

Frequency and conduction velocity analysis of the abductor pollicis brevis muscle during early fatigue

The physiological behavior of the abductor pollicis brevis (APB) muscle during early stage of fatigue is important as a reference for future clinical assessment of a pathologically altered muscle, as e.g. in carpal tunnel syndrome. The purpose of this study was to assess changes of force and surface electromyograms (sEMG) during early stage of fatigue of the APB. Thumb abduction force and sEMG derived from a multi-electrode array were recorded during isometric contraction. Electrode placement over the innervation zone (IZ) and the muscle tendon interface were avoided. The sEMGs of two adjacent electrode pairs were selected for the analysis, which yielded (a) motor unit conduction velocities (MUCV) derived from a correlation analysis between the EMGs and (b) mean frequencies obtained by using either fast Fourier (FMF) or Wavelet Transform (WMF). Early fatigue resulted in a relative decay rate of force (?2.1%¹ s^?1), MUCV (?1.5%¹ s^?1), FMF (?4.1%¹ s^?1), WMF (?3.7%¹ s^?1) and in a change of the power spectrum shape. Lower mean frequencies were observed at greater distances from the IZ independently of fatigue. The APB muscle seems to be fast fatigable and the relative decay rate of mean frequency was significantly larger than the one of force and MUCV.     

Concentric and eccentric isokinetic muscle activity separated by paired pattern classification of wavelet transformed mechanomyograms

It was hypothesized that concentric and eccentric isokinetic muscle actions should yield detectable differences in the mechanomyograms, which may reflect properties of the contraction and relaxation phases of the muscles. A paired pattern classification technique was adapted to determine whether wavelet transformed mechanomyograms from the three superficial quadriceps muscles were different during maximal concentric and eccentric isokinetic muscle actions. Mechanomyograms for this study were recorded from eleven healthy men (mean ± SD age = 20.1 ± 1.1 yrs) who performed maximal concentric and eccentric isokinetic muscle actions of the dominant leg extensors at a velocity of 30° s^?1. The results indicated that the paired pattern classification accurately classified the MMG intensity patterns in approximately 94% of the cases as being from a concentric or eccentric movement. Thus, it can be concluded that the differences in the intensity patterns recorded from concentric and eccentric muscle actions were significant. These findings indicated that the combined MMG wavelet analysis and pattern classification techniques could potentially be useful in situations where muscle activity during concentric muscle actions must be distinguished from that during eccentric muscle actions.