Structure of a helically extended SH3 domain of the T cell adapter protein ADAP

The adapter protein ADAP (FYB/SLAP-130) provides a critical link between T cell receptor (TCR) signaling and cell adhesion via the activation of integrins. The C-terminal 70 residues of ADAP show homology to SH3 domains; however, conserved residues of the fold are absent. An alignment and annotation of this domain has therefore been elusive. We have solved the three-dimensional structure of the ADAP C-terminal domain by NMR spectroscopy and show that it represents an altered SH3 domain fold. An N-terminal, amphipathic helix makes extensive contacts to residues of the regular SH3 domain fold, and thereby a composite surface with unusual surface properties is created. We propose this SH3 domain variant to be classified as a helically extended SH3 domain (hSH3 domain) and show that the ADAP-hSH3 domain can no longer bind conventional proline-rich peptides.     

Synthesis of a biotin-tagged photoaffinity probe of 2-azetidinone cholesterol absorption inhibitors

The design and synthesis of a biotin-tagged photoreactive analogue C-4 of the cholesterol absorption inhibitor Ezetimibe is described. Photoaffinity labeling of intestinal brush border membrane vesicles with C-4 and subsequent streptavidin-biotin chromatography leads to selective extraction of a 145 kDa integral membrane protein as the molecular target for cholesterol absorption inhibitors.     

Mutation in TECPR2 Reveals a Role for Autophagy in Hereditary Spastic Paraparesis

We studied five individuals from three Jewish Bukharian families affected by an apparently autosomal-recessive form of hereditary spastic paraparesis accompanied by severe intellectual disability, fluctuating central hypoventilation, gastresophageal reflux disease, wake apnea, areflexia, and unique dysmorphic features. Exome sequencing identified one homozygous variant shared among all affected individuals and absent in controls: a 1 bp frameshift TECPR2 deletion leading to a premature stop codon and predicting significant degradation of the protein. TECPR2 has been reported as a positive regulator of autophagy. We thus examined the autophagy-related fate of two key autophagic proteins, SQSTM1 (p62) and MAP1LC3B (LC3), in skin fibroblasts of an affected individual, as compared to a healthy control, and found that both protein levels were decreased and that there was a more pronounced decrease in the lipidated form of LC3 (LC3II). siRNA knockdown of TECPR2 showed similar changes, consistent with aberrant autophagy. Our results are strengthened by the fact that autophagy dysfunction has been implicated in a number of other neurodegenerative diseases. The discovered TECPR2 mutation implicates autophagy, a central intracellular mechanism, in spastic paraparesis.     

A method to obtain surface strains of soft tissues using a laser scanning device

A three-dimensional laser scanning device was developed allowing surface digitization of musculoskeletal and soft tissue structures under different loads. Image-processing algorithms were formulated for image registration. These were used to determine displacement mapping and then surface strains. Various validation experiments were performed. Accuracy was obtained on a test cylinder after rigid rotation and on a silicon cylinder compressed in four loading steps. The system accuracy (including the scanning and the data evaluation) was +/-0.10% strain in vertical and +/-0.16% strain in shear and circumferential direction for the rigid rotation exhibiting the zero-strain situation. Silicon cylinder compression showed that the accuracy was best for small strains, whereas strains >5% evoked a slight underestimation increasing further with higher strains (error of 0.54% for 7.22% vertical strain). It was possible to increase the accuracy by performing the strain measurements via sub-steps. This had a remaining error of 0.41% for 7.22% vertical strain. A further experiment was carried out in order to acquire the surface strain of a human lumbar intervertebral disc while it was forced to flexion and extension. This study introduced a laser-based scanning method to obtain soft tissue surface strains. It is important to know the strain distribution of musculoskeletal structures and soft tissues. This could help to better understand the mechanical loading of biological structures e.g. the processes in fracture healing. These data could also be used to assist in the validation process for finite-element models.     

Patchy distribution of flexible genetic elements in bacterial populations mediates robustness to environmental uncertainty

The generation and maintenance of genetic variation seems to be a general ecological strategy of bacterial populations. Thereby they gain robustness to irregular environmental change, which is primarily the result of the dynamic evolution of biotic interactions. A benefit of maintaining population heterogeneity is that only a fraction of the population has to bear the cost of not (yet) beneficial deviation. On evolutionary time frames, an added value of the underlying mechanisms is evolvability, i.e. the heritable ability of an evolutionary lineage to generate and maintain genetic variants that are potentially adaptive in the course of evolution. Horizontal gene transfer is an important mechanism that can lead to differences between individuals within bacterial populations. Broad host-range plasmids foster this heterogeneity because they are typically present in only a fraction of the population and provide individual cells with genetic modules newly acquired from other populations or species. We postulate that the benefit of robustness on population level could balance the cost of transfer and replication functions that plasmids impose on their hosts. Consequently, mechanisms that make a subpopulation conducive to specific conjugative plasmids may have evolved, which could explain the persistence of even cryptic plasmids that do not encode any traits.     

The cellular ceramide transport protein CERT promotes Chlamydia psittaci infection and controls bacterial sphingolipid uptake

Chlamydiaceae are bacterial pathogens that cause diverse diseases in humans and animals. Despite their broad host and tissue tropism, all Chlamydia species share an obligate intracellular cycle of development and have evolved sophisticated mechanisms to interact with their eukaryotic host cells. Here, we have analysed interactions of the zoonotic pathogen Chlamydia psittaci with a human epithelial cell line. We found that C. psittaci recruits the ceramide transport protein (CERT) to its inclusion. Chemical inhibition and CRISPR/Cas9‐mediated knockout of CERT showed that CERT is a crucial factor for C. psittaci infections thereby affecting different stages of the infection including inclusion growth and infectious progeny formation. Interestingly, the uptake of fluorescently labelled sphingolipids in bacteria inside the inclusion was accelerated in CERT‐knockout cells indicating that C. psittaci can exploit CERT‐independent sphingolipid uptake pathways. Moreover, the CERT‐specific inhibitor HPA‐12 strongly diminished sphingolipid transport to inclusions of infected CERT‐knockout cells, suggesting that other HPA‐12‐sensitive factors are involved in sphingolipid trafficking to C. psittaci. Further analysis is required to decipher these interactions and to understand their contributions to bacterial development, host range, tissue tropism, and disease outcome.     

Plasmonic Backscattering Effect in High‐Efficient Organic Photovoltaic Devices

A universal strategy for efficient light trapping through the incorporation of gold nanorods on the electron transport layer (rear) of organic photovoltaic devices is demonstrated. Utilizing the photons that are transmitted through the active layer of a bulk heterojunction photovoltaic device and would otherwise be lost, a significant enhancement in power conversion efficiency (PCE) of poly[N‐9′‐heptadecanyl‐2,7‐carbazole‐alt‐5,5‐(4′,7′‐di‐2‐thienyl‐2′,1′,3′‐benzothiadiazole)]:phenyl‐C₇₁‐butyric acid methyl ester (PCDTBT:PC₇₁BM) and poly[[4,8‐bis[(2‐ethylhexyl)oxy]benzo[1,2‐b:4,5‐b′]dithiophene‐2,6‐diyl][3‐fluoro‐2‐[(2‐ethylhexyl)carbonyl]thieno[3,4‐b] thiophenediyl]] (PTB7):PC₇₁BM by ≈13% and ≈8%, respectively. PCEs over 8% are reported for devices based on the PTB7:PC₇₁BM blend. A comprehensive optical and electrical characterization of our devices to clarify the influence of gold nanorods on exciton generation, dissociation, charge recombination, and transport inside the thin film devices is performed. By correlating the experimental data with detailed numerical simulations, the near‐field and far‐field scattering effects are separated of gold nanorods (Au NRs), and confidently attribute part of the performance enhancement to the enhanced absorption caused by backscattering. While, a secondary contribution from the Au NRs that partially protrude inside the active layer and exhibit strong near‐fields due to localized surface plasmon resonance effects is also observed but is minor in magnitude. Furthermore, another important contribution to the enhanced performance is electrical in nature and comes from the increased charge collection probability.     

Parietal area VIP neuronal responses to heading stimuli are encoded in head-centered coordinates

The ventral intraparietal area (VIP) is a multimodal parietal area, where visual responses are brisk, directional, and typically selective for complex optic flow patterns. VIP thus could provide signals useful for visual estimation of heading (self-motion direction). A central problem in heading estimation is how observers compensate for eye velocity, which distorts the retinal motion cues upon which perception depends. To find out if VIP could be useful for heading, we measured its responses to simulated trajectories, both with and without eye movements. Our results showed that most VIP neurons very strongly signal heading direction. Furthermore, the tuning of most VIP neurons was remarkably stable in the presence of eye movements. This stability was such that the population of VIP neurons represented heading very nearly in head-centered coordinates. This makes VIP the most robust source of such signals yet described, with properties ideal for supporting perception.