Background: Whether papain-like protease (PLpro) from avian infectious bronchitis virus (IBV) that infects poultry, causing huge losses, can perform deubiquitination is unknown. Results: IBV PLpro cleaves Lys 48 -and Lys 63 -linked polyubiquitin. The crystal structure reveals a markedly different set of residues for binding substrates. Conclusion: IBV PLpro performs deubiquitination with catalytic efficiencies that are different from other PLpros. Significance: Results provide a framework for developing antivirals.
Toona sinensis, popularly known as Chinese toon or Chinese mahogany, is a perennial deciduous arbor belonging to the genus Toona in the Meliaceae family, which is widely distributed and cultivated in eastern and southeastern Asia. Its fresh young leaves and buds have been consumed as a very popular nutritious vegetable in China and confirmed to display a wide variety of biological activities. To investigate the chemical constituents and their potential health benefits from the fresh young leaves and buds of T. sinensis, a phytochemical study on its fresh young leaves and buds was therefore undertaken. In our current investigation, 16 limonoids (1–16), including four new limonoids, toonasinenoids A–D (1–4), and a new naturally occurring limonoid, toonasinenoid E (5), were isolated and characterized from the fresh young leaves and buds of T. sinensis. The chemical structures and absolute configurations of limonoids 1–5 were elucidated by comprehensive spectroscopic data analyses. All known limonoids (6–16) were identified via comparing their experimental spectral data containing mass spectrometry data, 1H and 13C nuclear magnetic resonance data, and optical rotation values to the data reported in the literature. All known limonoids (6–16) were isolated from T. sinensis for the first time. Furthermore, the neuroprotective effects of all isolated limonoids 1–16 against 6-hydroxydopamine-induced cell death in human neuroblastoma SH-SY5Y cells were assessed in vitro. Limonoids 1–16 exhibited notable neuroprotective activities, with EC50 values in the range from 0.27 ± 0.03 to 17.28 ± 0.16 μM. These results suggest that regular consumption of the fresh young leaves and buds of T. sinensis might prevent the occurrence and development of Parkinson’s disease (PD). Moreover, the isolation and characterization of these limonoids that exhibit notable neuroprotective activities from the fresh young leaves and buds of T. sinensis could be very significant for researching and developing new neuroprotective drugs used for the prevention and treatment of PD.
Pseudomonas fluorescens GcM5-1A was isolated from the pine wood nematode (PWN), Bursaphelenchus xylophilus, obtained from wilted Japanese black pine, Pinus thumbergii, in China. In this paper, a genomic library of the GcM5-1A strain was constructed and a toxin-producing clone was isolated by bioassay. Nucleotide sequence analysis revealed an open reading frame of 1,290 bp encoding a protein of 429 amino acids with N-terminal putative signal peptide of 36 amino acids, which shared a similarity of 83, 82 and 80% identity with hypothetical protein PFLU2919 from P. fluorescens SBW25, Dyp-type peroxidase family protein from P. fluorescens Pf-5 and Tat-translocated enzyme from P. fluorescens Pf0-1, respectively. The gene encoding a full-length protein or without the putative signal peptide was cloned and expressed as a soluble protein in E. coli. The recombinant protein was purified to electrophoretic homogeneity by affinity chromatography using a Ni2+ matrix column. Its relative molecular weight was estimated to be 48.5 kDa by SDS-PAGE for full-length protein, and 45.0 kDa for the recombinant protein without putative signal peptide. Bioassay results showed that the recombinant protein with or without the putative signal peptide was toxic to both suspension cells and P. thunbergii seedlings. HPLC analysis demonstrated that components in branch extracts of P. thunbergii were significantly changed after addition of the recombinant full-length protein and hydrogen peroxide, which indicated that it is probably a peroxidase. This study offers information that can be used to determine the mechanism of pine wilt disease caused by the PWN.
Ti3C2 Tx nanosheet, the first synthesized MXene with high capacity performance and charge/discharge rate, has attracted increasingly attention in renewable energy storage applications. By performing systematic density functional theory calculations, the theoretical capacity of the intrinsic structure of single- and multi-layered Ti3C2 T 2 (T = F or O) corresponding to M (M = Li and Na) atoms are investigated. Theoretical volumetric capacity and gravimetric capacity are obtained, which are related to the stacking degree. The optimal ratios of capacity to structure are determined under different stacking degrees for understanding the influence of surface functional groups on energy storage performance. Its performance can be tuned by performing surface modification and increasing the interlayer distance. In addition, the reason for theoretical capacity differences of M atoms is analyzed, which is attributed to difference in interaction between the M-ions and substrate and the difference in electrostatic exclusion between adsorbed M-ions. These results provide an insight into the understanding of the method of efficiently increasing the energy storage performance, which will be useful for designing and using high performance electrode materials.
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