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Neospora caninum (NC) is a protozoan infection causing neosporosis, a disease that leads to substantial economic loss in livestock, especially in cattle, sheep, and goats. Unfortunately, there is presently no viable vaccination or authorized therapy available. Our research applied a bioinformatics approach to design a multi-epitope peptide (MEP) vaccine aimed at the pathogenic proteins GRA2 and Nc-p43 of NC, which are critical in parasite-mediated antigenicity and host interactions. Consequently, our research employs an in-silico methodology, including protein sequence retrieval, epitope prediction, vaccine design, structural analysis, molecular docking, molecular dynamics simulations, immunological simulation, and codon optimization using in silico cloning. We conducted analyses of antigenicity, allergenicity, toxicity, topology, and immunogenicity using multiple bioinformatics methods and identified 4 CTL, 4 HTL, and 2 B-cell epitopes. The vaccine design was created by integrating an adjuvant and a PADRE sequence to enhance immunogenicity, along with linkers (AAY, GPGPG, KK) to facilitate appropriate structural assembly, which was then analyzed for optimal complete profiles. Molecular docking with the Bos taurus (cattle) TLR9 receptor demonstrated a robust binding affinity, scoring − 1183.4, while molecular dynamics (MD) simulations over 50 ns validated persistent interactions between the vaccine and immune receptors. Moreover, immunological models forecasted a robust adaptive immune response, marked by an increase in the production of cytokines and the establishment of memory T-lymphocytes. Ultimately, codon optimization and in silico cloning validated the capacity for effective expression in E. coli . The results demonstrate that the developed vaccine has considerable immunogenic potential against NC. Nonetheless, more in vitro and in vivo studies are necessary to confirm its efficiency.

AIMP1/p43 protein is a structural component of multisynthetase complex (codosome) in eukaryotes, which reveals both tRNA binding and cytokine activities. Aim. Bacterial expression and purification of isotopically-labeled recombinant AIMP1/p43 protein in E. coli cells for studying its solution structure by multidimensional NMR spectroscopy. Methods. AIMP1/p43 protein was expressed in E. coli BL21(DE3)pLysE cells on M9 minimal medium with 15N isotope labeling and purified by metal-chelated chromatography. Heteronuclear 2D 1H-15N NMR experiments were performed in solution at 293 K on Agilent DDR2 800 NMR spectrometer. Results. The AIMP1/p43 protein was obtained in uniformly 15N-labeled form as an NMR sample. A high dispersion of resonance signals in the 2D 1H-15N HSQC NMR spectra confirmed the presence of its compact 3D protein structure. The NMR spectrum of AIMP1/p43 demonstrated a high signal-to-noise ratio and sufficient stability to acquire other multidimensional NMR data sets for determination of the structure of AIMP1/p43 protein in solution. Conclusions. The 15N-labeled AIMP1/p43 protein was stable for 4–7 days, which makes possible acquiring the critical NMR experimental data for detailed structural analysis in solution. Our data on the initial NMR spectra indicated the presence of some additional signals in comparison with the NMR spectrum of EMAP II which could be assigned to amino acids of the N-terminal α-helical fragment of AIMP1/p43.

Telomerase is the ribonucleoprotein enzyme responsible for the replication of chromosome ends in most eukaryotes. In the ciliate Euplotes aediculatus , the protein p43 biochemically co‐purifies with active telomerase and appears to be stoichiometric with both the RNA and the catalytic protein subunit of this telomerase complex. Here we describe cloning of the gene for p43 and present evidence that it is an authentic component of the telomerase holoenzyme. Comparison of the nucleotide sequence of the cloned gene with peptide sequences of the protein suggests that production of full‐length p43 relies on a programmed ribosomal frameshift, an extremely rare translational mechanism. Anti‐p43 antibodies immunodeplete telomerase RNA and telomerase activity from E.aediculatus nuclear extracts, indicating that the vast majority of mature telomerase complexes in the cell are associated with p43. The sequence of p43 reveals similarity to the La autoantigen, an RNA‐binding protein involved in maturation of RNA polymerase III transcripts, and recombinant p43 binds telomerase RNA in vitro . By analogy to other La proteins, p43 may function in chaperoning the assembly and/or facilitating nuclear retention of telomerase.

The localization of tyrosyl-tRNA synthetase has been studied by immunoelectronic microscopy in bovine kidney cells and fibroblasts of RAT I line which have been treated by monoclonal antibodies T3 raised against bovine tyrosyl-tRNA synthetase an by complexes of protein A-colloidal gold. The localization of tyrosyl-tRNA synthetase har been revealed both in cytoplasm and in the nucleus of mammalian cells. Tyrosyl-tRNA synthetase is located in cytoplasm mainly in the vicinity of polyribosomes what supports the compartmentatization conception of the components of protein synthesis apparatus. A significant portion of synthetase detected in the nucleus is located mainly in the region of diffuse chromatin, and partly in the nudeolus. In general, localization of tyrosyl-tRNA synthetase in mammalian cells is very similar to the localization of p43 protein of codosome, a precursor of the EMAP II cytokine, which is highly homologous to the non-catalytic C-terminal domain of tyrosyl-tRNA synthetase. Nuclear localization of tyrosyl-tRNA synthetase implies that this enzyme is involved in some non-canonical functions in the nucleus of eukaryotic cell. It is possible that this function may be related to the export of mature tRNA from nucleus to cytoplasm.

Trichuris whipworms cause disease and morbidity in humans and other animals. Their prolonged intestinal infections persist despite intact immune systems of their hosts and are attributed to immunomodulatory activities of their secretions. The p43 (Tm-DLP-1) protein of Trichuris muris of mice comprises 95% of the protein secreted by adult parasites, binds matrix proteoglycans, and has immune cytokine (IL-13)-neutralising activity. Using fluorescence-based methods we show that p43 binds fatty acids and retinol, including signalling lipids or precursors thereof. The orthologue of p43 from the human whipworm, Trichuris trichiura, exhibits similar lipid-binding activity. From the known molecular structure of p43, we explore the existence of extensive surface-accessible cavities with diverse surface charge characteristics which may indicate binding of diverse small molecule types, and its internally duplicated subdomains likely possess divergent characteristics. p43 represents a novel protein type (“dorylipophorin”) only known in Dorylaimia (Clade I) nematodes. We demonstrate that p43 is the dominant protein in Trichuris’s pseudocoelomic fluid, replacing the major internal lipid transporters of all other nematode clades, representing an ancient functional dichotomy. In Trichuris , and potentially other Clade I parasites of plants and animals, these proteins’ lipid-binding activities may be adapted for both internal physiological and external immunomodulatory activities.

Background Age-related hearing loss (ARHL), also known as presbycusis, is the most common sensory impairment seen in elderly people. However, the cochlear aging process does not affect people uniformly, suggesting that both genetic and environmental (e.g., noise, ototoxic drugs) factors and their interaction may influence the onset and severity of ARHL. Considering the potential links between thyroid hormone, mitochondrial activity, and hearing, here, we probed the role of p43, a N-terminally truncated and ligand-binding form of the nuclear receptor TRα1, in hearing function and in the maintenance of hearing during aging in p43 −/− mice through complementary approaches, including in vivo electrophysiological recording, ultrastructural assessments, biochemistry, and molecular biology. Results We found that the p43 −/− mice exhibit no obvious hearing loss in juvenile stages, but that these mice developed a premature, and more severe, ARHL resulting from the loss of cochlear sensory outer and inner hair cells and degeneration of spiral ganglion neurons. Exacerbated ARHL in p43 −/− mice was associated with the early occurrence of a drastic fall of SIRT1 expression, together with an imbalance between pro-apoptotic Bax, p53 expression, and anti-apoptotic Bcl2 expression, as well as an increase in mitochondrial dysfunction, oxidative stress, and inflammatory process. Finally, p43 −/− mice were also more vulnerable to noise-induced hearing loss. Conclusions These results demonstrate for the first time a requirement for p43 in the maintenance of hearing during aging and highlight the need to probe the potential link between human THRA gene polymorphisms and/or mutations and accelerated age-related deafness or some adult-onset syndromic deafness.

Aminoacyl-tRNA synthetase-interacting multifunctional protein 1 (AIMP1) is a non-catalytic component of the multi-tRNA synthetase complex which catalyzes the ligation of amino acids to the correct tRNAs. Pathogenic variants in several aminoacyl-tRNA synthetases genes have been linked to various neurological disorders, including leukodystrophies and pontocerebellar hypoplasias (PCH). To date, loss-of-function variants in AIMP1 have been associated with hypomyelinating leukodystrophy-3 (MIM 260600). Here, we report a novel frameshift AIMP1 homozygous variant (c.160delA,p.Lys54Asnfs) in a child with pontocerebellar hypoplasia and simplified gyral pattern, a phenotype not been previously described with AIMP1 variants, thus expanding the phenotypic spectrum. AIMP1 should be included in diagnostic PCH gene panels.

P43 is a truncated form of thyroid hormone receptor α localized in mitochondria, which stimulates mitochondrial respiratory chain activity. Previously, we showed that deletion of p43 led to reduction of pancreatic islet density and a loss of glucose-stimulated insulin secretion in adult mice. The present study was designed to determine whether p43 was involved in the processes of β cell development and maturation. We used neonatal, juvenile, and adult p43-/- mice, and we analyzed the development of β cells in the pancreas. Here, we show that p43 deletion affected only slightly β cell proliferation during the postnatal period. However, we found a dramatic fall in p43-/- mice of MafA expression (V-Maf Avian Musculoaponeurotic Fibrosarcoma Oncogene Homolog A), a key transcription factor of beta-cell maturation. Analysis of the expression of antioxidant enzymes in pancreatic islet and 4-hydroxynonenal (4-HNE) (a specific marker of lipid peroxidation) staining revealed that oxidative stress occurred in mice lacking p43. Lastly, administration of antioxidants cocktail to p43-/- pregnant mice restored a normal islet density but failed to ensure an insulin secretion in response to glucose. Our findings demonstrated that p43 drives the maturation of β cells via its induction of transcription factor MafA during the critical postnatal window.

p43 is a cofactor of aminoacyl-tRNA synthetase in mammals that effectively inhibits angiogenesis. However, the role of p43 in angiogenesis remains unclear. In the present study, we examined the effects of p43 on angiogenesis using human microvascular endothelial cells-1 (HMEC-1) cells as a model. Our microarray data showed that p43 regulated a number of cytokines, and the majoity of these are involved in the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway. IP-10 was previously shown to inhibit angiogenesis and suppress tumor growth via the JAK-STAT signaling pathway in vitro and in vivo. Our results showed that p43 induces both the mRNA and protein expression of IP-10. Furthermore, we demonstrated that p43 exerted an effect on the JAK-STAT signaling pathway by regulating key factors of the pathway. Using a JAK inhibitor, AG490, we studied the effect of p43 on HMEC-1 cells by blocking the JAK-STAT pathway. We found that AG490 inhibited the induction of IP-10 expression by p43, and suppressed the inhibitory effect of p43 on tubule formation and cell migration in HMEC-1 cells. We concluded that p43 inhibits tubule formation and cell migration by inducing IP-10 through the JAK-STAT signaling pathway, and blocking the JAK-STAT pathway with AG490 diminishes the inhibitory effects of p43 on angiogenesis.

Toxoplasma gondii is an obligate intracellular protozoan parasite, capable of infecting all species of mammals including man. Congenital toxoplasmosis is more important during pregnancy for the first time. In this study we expressed and purified P43 Toxoplasma gondii tachyzoite and bradyzoite specific surface antigen. The recombinant pGEMEX-1 contained Toxoplasma P43 coding sequence was transformed into E. coli and mass cultured in LB medium contained 100 μg/ml ampicillin at 37°C over night. The T7 promoter was induced by 1mM isopropyl-1-thio-ß-D-galactopyranoside (IPTG. Recombinant protein was purified by affinity chromatography and confirmed by gel diffusion dot blot and western blot,-using specific anti Toxoplasma antibodies. Recombinant plasmid was induced by IPTG and analyzed by SDS-PAGE. Recombinant protein was confirmed by Western-blot and dot blot using anti human Toxoplasma antibody. Recombinant Toxoplasma P43 was produced successfully.