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Feng Hong, Mengxue Zhao, Lei Zhang, Linyin Feng
Epigenetic modifications play an important role in neural development. Trimethylated histone H3 at lysine 27 (H3K27me3) is a repressive epigenetic mark that mediates tissue development. Here, we demonstrated that H3K27me3 and histone methyl-transferase Ezh2 regulated the development of dopaminergic neurons in vitro and in vivo. We found that H3K27me3 increased during ventral midbrain derived neural stem cells (VM-NSCs) differentiation. However, histone demethylase selective inhibitor GSK-J1 increased H3K27me3 level and decreased the expression of tyrosine hydroxylase (TH)...
March 19, 2019: Stem Cells and Development
Jenna Marion Gilkes, Campbell Sheen, Rebekah A Frampton, Grant R Smith, Renwick Dobson
'Candidatus Liberibacter solanacearum' (CLso) is an unculturable α-proteobacterium that is the causal agent of zebra chip disease of potato-a major problem in potato-growing areas, as it affects growth and yield. Developing effective treatments for CLso has been hampered by the difficulty in functionally characterizing the proteins of this organism, largely because they are not easily expressed and purified in standard expression systems. CLso has a reduced genome and its proteins are predicted to be prone to instability and aggregation...
March 19, 2019: Phytopathology
Lukas Schwintzer, Eva Aguado Roca, Meike Broemer
TRIAD3/RNF216 is a ubiquitin ligase of the RING-in-between-RING family. Recent publications identified TRIAD3 mutations in patients with neurological diseases, including Gordon Holmes syndrome and Huntington-like disorder. To understand the functional relevance of these disease-associated mutations, we have tested the ubiquitin ligase activity of mutated TRIAD3 in vitro. Several of these point mutations completely abrogated TRIAD3's catalytic activity. Using mass spectrometry, we identified new TRIAD3-interacting proteins/substrates from mouse brain lysate, which provide a new link between TRIAD3 and processes involving clathrin-mediated endocytosis...
2019: Cell Death Discovery
Ye Heng Lim, Hooi Ling Foo, Teck Chwen Loh, Rosfarizan Mohamad, Norhani Abdullah
Background: Increasing understanding on the functions of amino acids (AA) has led to new commercial applications and expansion of the worldwide markets. However, the current technologies rely heavily on non-food grade microorganism and chemical synthesis for the production of AA. Several studies reported that lactic acid bacteria (LAB) have the capability of producing AA owing to their well-established proteolytic system and amino acid biosynthesis genes. Hence, the objectives of this study were to explore the extracellular proteolytic activity of LAB isolated from various Malaysian fermented foods and their potential to produce AA extracellularly as feed supplements...
2019: Journal of Animal Science and Biotechnology
Takahiro Nakatani, Norihisa Yasui, Issei Tamura, Atsuko Yamashita
Green fluorescent protein (GFP) is amenable to recombinant expression in various kinds of cells and is widely used in life science research. We found that the recombinant expression of GFPuv, a commonly-used mutant of GFP, in E. coli produced two distinct molecular species as judged by in-gel fluorescence SDS-PAGE. These molecular species, namely form I and II, could be separately purified by anion-exchange chromatography without any remarkable differences in the fluorescence spectra. Mass spectrometric analyses revealed that the molecular mass of form I is almost the same as the calculated value, while that of form II is approximately 1 Da larger than that of form I...
March 18, 2019: Scientific Reports
Verjine Khodaverdian, Subhasish Tapadar, Ian A MacDonald, Yuan Xu, Po-Yi Ho, Allison Bridges, Pragya Rajpurohit, Bhakti A Sanghani, Yuhong Fan, Muthusamy Thangaraju, Nathaniel A Hathaway, Adegboyega K Oyelere
Deferiprone (DFP) is a hydroxypyridinone-derived iron chelator currently in clinical use for iron chelation therapy. DFP has also been known to elicit antiproliferative activities, yet the mechanism of this effect has remained elusive. We herein report that DFP chelates the Fe2+ ion at the active sites of selected iron-dependent histone lysine demethylases (KDMs), resulting in pan inhibition of a subfamily of KDMs. Specifically, DFP inhibits the demethylase activities of six KDMs - 2A, 2B, 5C, 6A, 7A and 7B - with low micromolar IC50 s while considerably less active or inactive against eleven KDMs - 1A, 3A, 3B, 4A-E, 5A, 5B and 6B...
March 18, 2019: Scientific Reports
Shanshan Liu, Weiqin Chang, Yuemei Jin, Chunyang Feng, Shuying Wu, Jiaxing He, Tianmin Xu
Cervical cancer is the fourth most common female cancer in the world. It is well known that cervical cancer is closely related to high-risk human papillomavirus (HPV) infection. However, epigenetics has increasingly been recognized for its role in tumorigenesis. Epigenetics refers to changes in gene expression levels based on non-gene sequence changes, primarily through transcription or translation of genes regulation, thus affecting its function and characteristics. Typical post-translational modifications (PTM) include acetylation, propionylation, butyrylation, malonylation and succinylation, among which the acetylation modification of lysine sites has been studied more clearly so far...
March 18, 2019: Bioscience Reports
Xuhao Ni, Wei Kou, Jian Gu, Ping Wei, Xiao Wu, Hao Peng, Jinhui Tao, Wei Yan, Xiaoping Yang, Andriana Lebid, Benjamin V Park, Zuojia Chen, Yizhu Tian, Juan Fu, Stephanie Newman, Xiaoming Wang, Hongbin Shen, Bin Li, Bruce R Blazar, Xuehao Wang, Joseph Barbi, Fan Pan, Ling Lu
Regulatory T cells (Tregs) are crucial mediators of immune control. The characteristic gene expression and suppressive functions of Tregs depend considerably on the stable expression and activity of the transcription factor FOXP3. Transcriptional regulation of the Foxp3 gene has been studied in depth, but both the expression and function of this factor are also modulated at the protein level. However, the molecular players involved in posttranslational FOXP3 regulation are just beginning to be elucidated. Here, we found that TRAF6-deficient Tregs were dysfunctional in vivo ; mice with Treg-restricted deletion of TRAF6 were resistant to implanted tumors and displayed enhanced anti-tumor immunity...
March 18, 2019: EMBO Journal
Amjad Ali, Sabihur Rahman Farooqui, Akhil C Banerjea
HIV-1 Tat is degraded in the host cell both by proteasomal and lysosomal pathways, but the specific molecules that engage with Tat from these pathways are not known. Since E3 ubiquitin ligases are the primary determinants of substrate specificity within the ubiquitin dependent proteasomal degradation of proteins, we first sought to identify the E3 ligase associated with Tat degradation. Based on the intrinsic disordered nature of Tat protein we focused our attention on host cell E3 ubiquitin ligase CHIP (carboxy terminus of Hsp70-binding protein)...
March 18, 2019: Journal of Biological Chemistry
Michael J Howard, K Grace Foley, David D Shock, Vinod K Batra, Samuel H Wilson
DNA methylation is an epigenetic mark that regulates gene expression in mammals. One method of methylation removal is through TET-catalyzed oxidation and the base excision repair pathway. The iterative oxidation of 5-methylcytosine (5mC) catalyzed by TET enzymes produces three oxidized forms of cytosine: 5-hydroxmethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxycytosine (5caC). The effect these modifications have on the efficiency and fidelity of the base excision repair pathway during the repair of opposing base damage, and in particular DNA polymerization, remains to be elucidated...
March 18, 2019: Journal of Biological Chemistry
Sara Klee, Judith P Sinn, Aleah Holmes, Brian L Lehman, Teresa Krawczyk, Kari A Peter, Timothy W McNellis
Elongation factor P (EF-P) facilitates the translation of certain peptide motifs, including those with multiple proline residues. EF-P must be post-translationally modified for full functionality; in Enterobacteria this is accomplished by two enzymes, EpmA and EpmB, which catalyze the β-lysylation of EF-P at a conserved lysine position. Mutations to efp or its modifying enzymes produce pleiotropic phenotypes, including decreases in virulence, swimming motility, and extracellular polysaccharide production, as well as proteomic perturbations...
March 18, 2019: Journal of Bacteriology
Jannel Acosta, Francisco Roa, Iván González-Chavarría, Allison Astuya, Rafael Maura, Raquel Montesino, Carolina Muñoz, Frank Camacho, Paulina Saavedra, Ariel Valenzuela, Oliberto Sánchez, Jorge R Toledo
Antimicrobial peptides (AMPs) are amphipathic peptides, which play an important role in innate defence. These peptides are gene-encoded and either constitutively expressed and/or upregulated during an infection. NK-lysins are AMPs with a three-dimensional globular structure. They are larger molecules, which comprise 74-78 amino acid residues and six conserved cysteine residues forming three disulphide bonds. Cathelicidins are a family of antimicrobial peptides that act as important components of the innate immune system with a broad spectrum of antimicrobial activity and immunomodulatory properties...
March 15, 2019: Fish & Shellfish Immunology
Moritz Beck-Broichsitter
Despite the significant interest in therapeutic antibodies for the treatment of airway diseases, no study addressed the challenge, which can arise when such formulations need to be made accessible for nebulization in concentrated (viscous) form. By 1) determining the maximum viscosity, which can still be atomized by vibrating-mesh technology and 2) supplementing the antibody formulation under investigation with at least one excipient, which decreases the viscosity under that specific threshold value of the utilized inhaler (and maintains the stability of the formulation), it should be possible to nebulize concentrated antibody formulations...
March 15, 2019: Journal of Pharmaceutical Sciences
Lu Cao, Xiao Man Wu, Pin Nie, Ming Xian Chang
CD44 gene is a cell surface receptor which undergoes complex alternative splicing and extensive post-translational modifications. Although many studies have showed that CD44 is involved in the process of host defense, the function of piscine CD44 in antibacterial or antiviral defense response remains unclear. In the present study, we report the functional characterization of zebrafish CD44c, which is more similar to CD44b antigen isoforms rather than CD44a based on amino acid composition and phylogenetic analysis...
March 15, 2019: Developmental and Comparative Immunology
Priyanga Ranasinghe, Ranil Jayawardena, Lal Chandrasena, Vivianne Noetzel, John Burd
BACKGROUND: Diabetes mellitus is rapidly becoming one of the leading causes of morbidity and mortality worldwide. Preventive measures have become important, especially in the South Asian region and other parts of the world where diabetes is becoming increasingly prevalent. We postulate that a product containing amino acid lysine, micronutrient zinc and vitamin C will have beneficial effects on glycemic control and disease progression in patients with pre-diabetes and we aim to evaluate this hypothesis using a phase II/III randomized double-blind controlled clinical trial design...
March 18, 2019: Trials
Marion Wobser, Alexandra Weber, Amelie Glunz, Saskia Tauch, Kristina Seitz, Tobias Butelmann, Sonja Hesbacher, Matthias Goebeler, René Bartz, Hella Kohlhof, David Schrama, Roland Houben
BACKGROUND: Targeting epigenetic modifiers is effective in cutaneous T cell lymphoma (CTCL). However, there is a need for further improvement of this therapeutic approach. Here, we compared the mode of action of romidepsin (FK228), an established class I histone deacetylase inhibitor, and domatinostat (4SC-202), a novel inhibitor of class I HDACs, which has been reported to also target the lysine-specific histone demethylase 1A (LSD1). METHODS: We performed MTS assays and flow cytometric analyses of propidium iodide or annexin V-stained cells to assess drug impact on cellular proliferation, cell cycle distribution, and survival...
March 18, 2019: Journal of Hematology & Oncology
Myriam Merarchi, Gautam Sethi, Muthu K Shanmugam, Lu Fan, Frank Arfuso, Kwang Seok Ahn
Histone deacetylases (HDACs) are enzymes that can control transcription by modifying chromatin conformation, molecular interactions between the DNA and the proteins as well as the histone tail, through the catalysis of the acetyl functional sites removal of proteins from the lysine residues. Also, HDACs have been implicated in the post transcriptional process through the regulation of the proteins acetylation, and it has been found that HDAC inhibitors (HDACi) constitute a promising class of pharmacological drugs to treat various chronic diseases, including cancer...
March 16, 2019: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Francisco Procura, Dante J Bueno, Susana B Bruno, Ariel D Rogé
This study was conducted to estimate the apparent prevalence of Salmonella spp. in chicken livers obtained from markets in Entre Ríos, Argentina, using two culture methods (preenrichment and direct selective agar plating). We also determined the antimicrobial resistance of Salmonella isolated strains and evaluated the performance of the two culture methods and selective-differential plating media used for Salmonella isolation. Of 666 chicken livers studied, 32 organs (4.8%) related to 4 poultry slaughterhouse companies were positive for Salmonella sp...
May 2019: Food Research International
Corrie Landymore, Timothy D Durance, Anika Singh, Anubhav Pratap Singh, David D Kitts
Krill, (Euphausia pacifica) contains a high protein content (>15.4%) and an estimated biological value higher than many animal protein sources. Thus it is considered to be an important source of high-quality protein. However, commercial processing of krill is limited due to problems such as presence of hydrolytic enzymes (proteases, carboxypeptidases, nucleases, and phospholipases), and its small size. These enzymes are released immediately upon krill harvesting, resulting in autolysis, and rapid spoilage...
May 2019: Food Research International
Ivan Ding, Jenna A Walz, Charles R Mace, Amy M Peterson
Polyelectrolyte multilayers (PEMs) are a category of materials commonly used as coatings on surfaces that interact with cells. The properties of PEMs have been established to be controlled by not only polyelectrolyte choice, but by the identity of the initially applied (bottom) layer. In this work, 5-bilayer PEMs consisting of poly(diallyldimethylammonium chloride) (PDADMAC) and poly(sodium 4-styrenesulfonate) (PSS) were coated on gold-sputtered quartz substrates with different first layer materials. A final poly-l-lysine (PLL) layer was added to all PEMs to provide identical top layers conducive to cell growth...
February 28, 2019: Colloids and Surfaces. B, Biointerfaces
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