Structural Adaptation of the Single-Stranded DNA-Binding Protein C-Terminal to DNA Metabolizing Partners Guides Inhibitor Design

Single-stranded DNA-binding protein (SSB) is a bacterial interaction hub and an appealing target for antimicrobial therapy. Understanding the structural adaptation of the disordered SSB C-terminus (SSB-Ct) to DNA metabolizing enzymes (e.g., ExoI and RecO) is essential for designing high-affinity SSB mimetic inhibitors. Molecular dynamics simulations revealed the transient interactions of SSB-Ct with two hot spots on ExoI and RecO. The residual flexibility of the peptide–protein complexes allows adaptive molecular recognition. Scanning with non-canonical amino acids revealed that modifications at both termini of SSB-Ct could increase the affinity, supporting the two-hot-spot binding model. Combining unnatural amino acid substitutions on both segments of the peptide resulted in enthalpy-enhanced affinity, accompanied by enthalpy–entropy compensation, as determined by isothermal calorimetry. NMR data and molecular modeling confirmed the reduced flexibility of the improved affinity complexes. Our results highlight that the SSB-Ct mimetics bind to the DNA metabolizing targets through the hot spots, interacting with both of segments of the ligands

Cell Delivery: Routing Nanomolar Protein Cargoes to Lipid Raft-Mediated/Caveolar Endocytosis through a Ganglioside GM1-Specific Recognition Tag (Adv. Sci. 4/2020)

In article number 1902621, Tamás A. Martinek and co-workers develop a pentapeptidic tag, which reads the glycan code of ganglioside GM1 and triggers lipid raft-mediated endocytosis, avoiding lysosomal entrapment. This carrier molecule can deliver macromolecular cargoes (e.g., IgG complexes) into live cells with the possibility to escape to the cytosol.Peer reviewe

Routing Nanomolar Protein Cargoes to Lipid Raft-Mediated/Caveolar Endocytosis through a Ganglioside GM1‐Specific Recognition Tag

There is a pressing need to develop ways to deliver therapeutic macromolecules to their intracellular targets. Certain viral and bacterial proteins are readily internalized in functional form through lipid raft-mediated/caveolar endocytosis, but mimicking this process with protein cargoes at therapeutically relevant concentrations is a great challenge. Targeting ganglioside GM1 in the caveolar pits triggers endocytosis. A pentapeptide sequence WYKYW is presented, which specifically captures the glycan moiety of GM1 (K-D = 24 nm). The WYKYW-tag facilitates the GM1-dependent endocytosis of proteins in which the cargo-loaded caveosomes do not fuse with lysosomes. A structurally intact immunoglobulin G complex (580 kDa) is successfully delivered into live HeLa cells at extracellular concentrations ranging from 20 to 160 nm, and escape of the cargo proteins to the cytosol is observed. The short peptidic WYKYW-tag is an advantageous endocytosis routing sequence for lipid raft-mediated/caveolar cell delivery of therapeutic macromolecules, especially for cancer cells that overexpress GM1.Peer reviewe

Individual and combined effects of feed artificially contaminated with with fumonisin B 1 and T-2 toxin in weaned rabbits

Co-contamination of feed and feed raw materials with two or more mycotoxins is frequently reported, however, only a few studies have investigated the combined effects of low doses of multiple mycotoxins. In the present study the individual and combined effects of 10 mg/kg fumonisin B1 and 2 mg/kg T-2 toxin (n=12/group) were investigated in weaned rabbits. Mycotoxin contaminated feed was produced by adding fungal cultures of Fusarium verticillioides and Fusarium sporotrichioides, and fed to 40 days old rabbits during 28 days. Feed intake and body weight were measured weekly, serum biochemistry and antioxidant parameters on day 0, 14 and 28, while histopathological examination and comet assay were performed at the end of the experiment. T-2 exposure both alone and in combination resulted in 15-18% less final body weight compared to the control and FB1 treatment. There was a significant increase in the concentration of plasma total protein, albumin, fructosamine and creatinine in the group treated with FB1 compared to the control. The liver and the kidney of most animals treated with T-2 toxin, FB1 and their combination showed pathological changes, occurring more frequent in animals exposed to both toxins. T-2 resulted in depletion of lymphocytes in the spleen. FB1 and T-2 exerted synergistic effect on the antioxidant/oxidative parameters after 2 weeks of exposure, manifesting in less glutathione and glutathione peroxidase, while more malondialdehyde was produced. Both toxins caused DNA damage in the lymphocytes, which was more pronounced in the group fed T-2 toxin and T-2 combined with FB1, without additive or synergistic effects

Synthesis and Biological Evaluation of Triazolyl 13α-Estrone–Nucleoside Bioconjugates

2′-Deoxynucleoside conjugates of 13α-estrone were synthesized by applying the copper-catalyzed alkyne–azide click reaction (CuAAC). For the introduction of the azido group the 5′-position of the nucleosides and a propargyl ether functional group on the 3-hydroxy group of 13α-estrone were chosen. The best yields were realized in our hands when the 3′-hydroxy groups of the nucleosides were protected by acetyl groups and the 5′-hydroxy groups were modified by the tosyl–azide exchange method. The commonly used conditions for click reaction between the protected-5′-azidonucleosides and the steroid alkyne was slightly modified by using 1.5 equivalent of Cu(I) catalyst. All the prepared conjugates were evaluated in vitro by means of MTT assays for antiproliferative activity against a panel of human adherent cell lines (HeLa, MCF-7 and A2780) and the potential inhibitory activity of the new conjugates on human 17β-hydroxysteroid dehydrogenase 1 (17β-HSD1) was investigated via in vitro radiosubstrate incubation. Some protected conjugates displayed moderate antiproliferative properties against a panel of human adherent cancer cell lines (the protected cytidine conjugate proved to be the most potent with IC50 value of 9 μM). The thymidine conjugate displayed considerable 17β-HSD1 inhibitory activity (IC50 = 19 μM)

A cigaretta csomagolás, design, stílus hatása fogyasztókra

A cigaretták csomagolásának hatását fogyasztókra mutatja be a csomagolás, design, stílus szempontjából. A Philip Morris Magyarország Kft. termékein keresztül értékeli, mutatja be és elemzi a különböző hatásokat. A fogyasztókon keresztül a vállalatot érő pozitív és negatív hatás is hangot kapott.Bav

Phytoplasma infection status survey in plum psyllid (Cacopsylla pruni) population

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