Who with whom: functional coordination of E2 enzymes by RING E3 ligases during poly-ubiquitylation

Protein modification with poly-ubiquitin chains is a crucial process involved in a myriad of cellular pathways. Chain synthesis requires two steps: substrate modification with ubiquitin (priming) followed by repetitive ubiquitin-to-ubiquitin attachment (elongation). RING-type E3 ligases catalyze both reactions in collaboration with specific priming and elongating E2 enzymes. We provide kinetic insight into poly-ubiquitylation during protein quality control by showing that priming is the rate-determining step in protein degradation as directed by the yeast ERAD RING E3 ligases, Hrd1 and Doa10. Doa10 cooperates with the dedicated priming E2, Ubc6, while both E3s use Ubc7 for elongation. Here, we provide direct evidence that Hrd1 uses Ubc7 also for priming. We found that Ubc6 has an unusually high basal activity that does not require strong stimulation from an E3. Doa10 exploits this property to pair with Ubc6 over Ubc7 during priming. Our work not only illuminates the mechanisms of specific E2/E3 interplay in ERAD, but also offers a basis to understand how RING E3s may have properties that are tailored to pair with their preferred E2s

Structural basis of glycan276-dependent recognition by HIV-1 broadly neutralizing antibodies

Recognition of N-linked glycan at residue N276 (glycan276) at the periphery of the CD4-binding site (CD4bs) on the HIV-envelope trimer is a formidable challenge for many CD4bs-directed antibodies. To understand how this glycan can be recognized, here we isolate two lineages of glycan276-dependent CD4bs antibodies. Antibody CH540-VRC40.01 (named for donor-lineage.clone) neutralizes 81% of a panel of 208 diverse strains, while antibody CH314-VRC33.01 neutralizes 45%. Cryo-electron microscopy (cryo-EM) structures of these two antibodies and 179NC75, a previously identified glycan276-dependent CD4bs antibody, in complex with HIV-envelope trimer reveal substantially different modes of glycan276 recognition. Despite these differences, binding of glycan276-dependent antibodies maintains a glycan276 conformation similar to that observed in the absence of glycan276-binding antibodies. By contrast, glycan276-independent CD4bs antibodies, such as VRC01, displace glycan276 upon binding. These results provide a foundation for understanding antibody recognition of glycan276 and suggest its presence may be crucial for priming immunogens seeking to initiate broad CD4bs recognition

Selenium nanoparticles synthesized using an eco-friendly method: Dye decolorization from aqueous solutions, cell viability, antioxidant, and antibacterial effectiveness

International audienceSelenium nanoparticles (SeNPs) were fabricated using a green microwave technique in the presence of ascorbic acid. The morphological features indicated that the semi-spherical SeNPs with a diameter 8.5-22nm were configured in agglomerated spherical shapes with diameters around 0.47-0.71 μm. Furthermore, the removal of Fuchsin Basic dye from aqueous solutions was investigated upon variation of concentration of SeNPs. The degradation efficiency achieved 100 % for 10 mg of SeNPs after 34 min of visible light irradiation time. The antioxidant activity 2 was tested via DPPH radical scavenging assay and displayed that the highest scavenging capacity (311.115.72 mg/g) was achieved by SeNPs at a concentration of 106.25 mg/mL. Otherwise, the cell viability of SeNPs through human fibroblasts cell lines in-vitro was reduced to be 75.13.8 % with nanoparticle concentration around 500 μg/mL. The antibacterial activity was investigated against gram-negative and gram-positive bacteria such as Escherichia coli (E.coli), Pseudomonas aeruginosa (P. aeruginosa), Klebsiella pneumoniae (K. pneumonia), Staphylococcus aureus (S. aureus), and Bacillus subtilis (B. subtilis) bacteria after one day of exposure. It was illustrated that SeNPs did not display an activity towards Staphylococcus aureus, while it possessed the highest one against Escherichia coli with MBC of 50 ± 1.76 g/mL compared with 26 ± 0.6 g/mL for the standard antibiotic. These tremendous properties of SeNPs indicate that manipulating multifunctional nanoparticles for versatile wound and skin treatment applications is highly encouraging

Top-down synthesis of graphene: A comprehensive review

Graphene research has become an emerging frontier in materials science because of its potential as a versatile material in multiple applications, from electronics, sensors, water treatment, batteries, displays, advanced composites, and coatings to biomedical applications. While the community has witnessed tremendous advances in the laboratory-scale synthesis of graphene, it is crucial to focus on sustainable large-scale graphene production to adopt graphene-based technology at an industrial scale. Several top-down and bottom-up methods have been developed to realize affordable graphene production. However, a low-cost scalable graphene production method with acceptable quality remains a challenge; top-down processes are demonstrating their potential to offer a more straightforward solution. Herein, we present an overview of recent progress in the research and development of top-down graphene synthesis methods and their potential to scale-up graphene production. We cover the effect of different synthesis parameters on the quality control of graphene. In addition, we provide a brief overview of bottom-up methods. Finally, we discuss the existing challenges and future directions in top-down methods for large-scale graphene production