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Biochemical Journal

Teresa M Buck, Alexa S Jordahl, Megan E Yates, Mike Preston, Emily Cook, Thomas R Kleyman, Jeffrey L Brodsky
The Epithelial Sodium Channel (ENaC) in kidney regulates blood pressure through control of sodium and volume homeostasis, and in lung ENaC regulates the volume of airway and alveolar fluids.  ENaC is a heterotrimer of homologous α-, β-, and γ-subunits, and assembles in the Endoplasmic Reticulum (ER) before it traffics and functions at the plasma membrane. Improperly folded or orphaned ENaC subunits are subject to ER quality control and targeted for ER associated degradation (ERAD). We previously established that a conserved, ER lumenal, molecular chaperone, Lhs1/GRP170, selects αENaC, but not β- or γENaC, for degradation when the ENaC subunits were individually expressed...
November 30, 2016: Biochemical Journal
Ryan P Emptage, Mark A Lemmon, Kathryn M Ferguson
Protein kinases are frequently regulated by intramolecular autoinhibitory interactions between protein modules that are reversed when these modules bind other 'activating' protein or membrane-bound targets.  One group of kinases, the M AP/ m icrotubule a ffinity- r egulating k inases (MARKs) contain a poorly understood regulatory module, the KA1 ( k inase a ssociated 1) domain, at their C-terminus.  KA1 domains from MARK1 and several related kinases from yeast to humans have been shown to bind membranes containing anionic phospholipids, and peptide ligands have also been reported...
November 22, 2016: Biochemical Journal
Catherine L Shelton, Deborah G Conrady, Andrew B Herr
Staphylococcus epidermidis is an opportunistic pathogen that can form robust biofilms that render the bacteria resistant to antibiotic action and immune responses. Intercellular adhesion in S. epidermidis biofilms is mediated by the cell wall-associated Accumulation-Associated Protein (Aap), via zinc-mediated self-assembly of its B-repeat region. This region contains up to 17 nearly-identical sequence repeats, with each repeat assumed to be functionally equivalent. However, Aap B-repeats exist as two subtypes, defined by a cluster of consensus or variant amino acids...
November 21, 2016: Biochemical Journal
Tetsu Shimizu, Lulu Yin, Ayako Yoshida, Yuusuke Yokooji, Shin-Ichi Hachisuka, Takaaki Sato, Takeo Tomita, Hiromi Nishida, Haruyuki Atomi, Tomohisa Kuzuyama, Makoto Nishiyama
β-Decarboxylating dehydrogenases, which are involved in central metabolism, are considered to have diverged from a common ancestor with broad substrate specificity. In a molecular phylogenetic analysis of 183 β-decarboxylating dehydrogenase homologs from 84 species, TK0280 from Thermococcus kodakarensis was selected as a candidate for an ancestral-type β-decarboxylating dehydrogenase. The biochemical characterization of recombinant TK0280 revealed that the enzyme exhibited dehydrogenase activities toward homoisocitrate, isocitrate, and 3-isopropylmalate, which correspond to key reactions involved in the lysine biosynthetic pathway, tricarboxylic acid cycle, and leucine biosynthetic pathway, respectively...
November 9, 2016: Biochemical Journal
Pablo N Romasanta, Lucrecia M Curto, María Belen Sarratea, Sofía Noly Truant, María Belen Antonoglou, María J Fernández Lynch, Jose María Delfino, Roy A Mariuzza, Marisa M Fernández, Emilio L Malchiodi
Natural Killer (NK) cells are lymphocytes of the innate immune system that eliminate virally infected or malignantly transformed cells. NK cell function is regulated by diverse surface receptors that are both activating and inhibitory. Among them, the homodimeric Ly49 receptors control NK cell cytotoxicity by sensing MHC class I molecules on target cells. Although crystal structures have been reported for Ly49/MHC-I complexes, the underlying binding mechanism has not been elucidated. Accordingly, we carried out thermodynamic and kinetic experiments on the interaction of four Ly49 receptors (Ly49G, Ly49H, Ly49I and Ly49P) with two MHC-I ligands (H-2D(d) and H-2D(k))...
November 9, 2016: Biochemical Journal
Iman Momken, Adrian Chabowski, Ellen Dirkx, Miranda Nabben, Swati Jain, Jan Glatz, Joost Luiken, Arend Bonen
Leptin stimulates fatty acid oxidation in muscle and heart; but, the mechanism by which these tissues provide additional intracellular fatty acids for their oxidation remains unknown. We examined, in isolated muscle and cardiac myocytes, whether leptin, via AMPK activation, stimulated FAT/CD36-mediated fatty acid uptake to enhance fatty acid oxidation. In both mouse skeletal muscle and rat cardiomyocytes, leptin increased fatty acid oxidation, an effect that was blocked when AMPK phosphorylation was inhibited by Ara-A or Compound C...
November 8, 2016: Biochemical Journal
Dakshnamurthy Selvakumar, Marian J Drescher, Nathan A Deckard, Neeliyath A Ramakrishnan, Barbara J Morley, Dennis G Drescher
Dopamine receptors regulate exocytosis via protein-protein interactions (PPIs) as well as via adenylyl cyclase transduction pathways. Evidence has been obtained for PPIs in inner ear hair cells coupling D1A to SNARE-related proteins snapin, otoferlin, NSF, and AP2mu1, dependent on [Ca(2+)] and phosphorylation. Specifically, the carboxy terminus of dopamine D1A was found to directly bind t-SNARE-associated protein snapin in teleost and mammalian hair cell models by yeast two-hybrid and pull-down assays, and snapin directly interacts with  hair cell calcium-sensor otoferlin...
November 7, 2016: Biochemical Journal
Florent Colomb, Marie-Ange Krzewinski-Recchi, Agata Steenackers, Audrey Vincent, Anne Harduin-Lepers, Philippe Delannoy, Sophie Groux-Degroote
We have previously shown that TNF induced the up-regulation of the sialyltransferase gene ST3GAL4 BX transcript through mitogen- and stress-activated kinase 1/2 (MSK1/2), extracellular-signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) signaling pathways. This up-regulation resulted in sialyl-Lewis(x) (sLe(x)) over-expression on high-molecular weight glycoproteins in inflamed airway epithelium and increased the adhesion of Pseudomonas aeruginosa PAO1 and PAK strains to lung epithelial cells...
November 7, 2016: Biochemical Journal
Natalie Gumataotao, K P Wasantha Lankathilaka, Brian Bennett, Richard C Holz
The Fe-type nitrile hydratase activator protein from Rhodococcus equi TG328-2 ( Re NHase-TG328-2) was successfully expressed and purified.  Sequence analysis and homology modeling suggest that it is a G3E P-loop GTPase within the COG0523 subfamily.  Kinetic studies revealed that the Fe-type activator protein is capable of hydrolyzing GTP to GDP with a k cat value of 1.2 x 10(-3) s(-1) and a K m value of 40 μM in the presence of 5 mM MgCl2 in 50 mM HEPES at pH 8.0.  The addition of divalent metal ions such as Co(II), which binds to the Re NHase-TG328-2 activator protein with a K d of 2...
November 2, 2016: Biochemical Journal
Joern Krausze, Corinna Probst, Ute Curth, Joachim Reichelt, Sayantan Saha, David Schafflick, Dirk Werner Heinz, Ralf Rainer Mendel, Tobias Kruse
The molybdenum cofactor (Moco) is a redox active prosthetic group, essentially required for numerous enzyme-catalyzed two electron transfer reactions. Moco is synthesized by an evolutionary old and highly conserved multi-step pathway. In the last step of Moco biosynthesis the molybdenum centre is inserted into the final Moco precursor adenylated molybdopterin (MPT-AMP). This unique and yet poorly characterized maturation reaction finally yields physiologically active Moco. In the model plant Arabidopsis , the two domain enzyme Cnx1 is required for Moco formation...
November 1, 2016: Biochemical Journal
Almudena Val-Blasco, Patricia Prieto-Chinchilla, Silvia Gonzalez-Ramos, Gemma Benito, María Teresa Vallejo-Cremades, Ivette Pacheco, María Pilar González-Peramato, Noelia Agra, Verónica Terrón, Carmen Delgado, Paloma Martín-Sanz, Lisardo Boscá, María Fernández-Velasco
Cardiac fibrosis and chronic inflammation are common complications in type 2 diabetes mellitus (T2D). Since nucleotide oligomerization-binding domain 1 (NOD1), an innate immune receptor, is involved in the pathogenesis of insulin resistance and diabetes outcomes, we sought to investigate its involvement in cardiac fibrosis. Here we show that selective staining of cardiac fibroblasts from T2D ( db/db;db ) mice exhibit up-regulation and activation of the NOD1 pathway, resulting in enhanced NF-kB and TGF-β signalling...
November 1, 2016: Biochemical Journal
Keisuke Obara, Akio Kihara
Yeast cells sense alterations in the plasma membrane (PM) lipid asymmetry and external alkalization by the sensor protein Rim21, which functions in the Rim101 pathway. Rim101 signaling is initiated at the PM by the recruitment of the Rim101 signaling complex. The PM physically associates with the cortical endoplasmic reticulum (ER) to form ER-PM contact sites, where several signaling events, lipid exchange, and ion transport take place. In the present study, we investigated the spatial relationship between ER-PM contact sites and the sites of Rim101 signaling...
November 1, 2016: Biochemical Journal
Ankit Srivastava, Sakshi Sharma, Sandhya Sadanandan, Sakshi Gupta, Jasdeep Singh, Sarika Gupta, V Haridas, Bishwajit Kundu
Misfolding and aggregation of cellular prion protein (PrP(C)) is associated with a large array of neurological disorders commonly called the Transmissible spongiform encephalopathies (TSEs). Designing inhibitors against prions has remained a daunting task owing to limited information about mechanism(s) of their pathogenic self-assembly. Here, we explore the anti-prion properties of a combinatorial library of bispidine-based peptidomimetics (BPMs) that conjugate amino acids with hydrophobic and aromatic side chains...
November 1, 2016: Biochemical Journal
Helen R Heathcote, Sarah J Mancini, Anastasiya Strembitska, Kunzah Jamal, James A Reihill, Timothy M Palmer, Gwyn W Gould, Ian P Salt
The key metabolic regulator, AMP-activated protein kinase (AMPK) is reported to be downregulated in metabolic disorders, but the mechanisms are poorly characterised. Recent studies have identified phosphorylation of the AMPKα1/α2 catalytic subunit isoforms at Ser487/491 respectively as an inhibitory regulation mechanism. Vascular endothelial growth factor (VEGF) stimulates AMPK and protein kinase B (Akt) in cultured human endothelial cells. As Akt has been demonstrated to be an AMPKα1 Ser487 kinase, the effect of VEGF on inhibitory AMPK phosphorylation in cultured primary human endothelial cells was examined...
October 26, 2016: Biochemical Journal
Enrico Malito, Paola Lo Surdo, Daniele Veggi, Laura Santini, Heather Heather Stefek, Brunella Brunelli, Enrico Luzzi, Matthew James Bottomley, Peter Beernink, Maria Scarselli
Factor H binding protein (fHbp) is an important antigen of Neisseria meningitidis that is able to elicit a robust protective immune response in humans. Previous studies on the interactions of fHbp with antibodies revealed that some anti-fHbp monoclonal antibodies that are unable to trigger complement-mediated bacterial killing in vitro , are highly cooperative and become bactericidal if used in combination. Several factors have been shown to influence such cooperativity, including IgG subclass and antigen density...
October 26, 2016: Biochemical Journal
Nidhi Kundu, Swapnil Tichkule, Shashi Bhushan Pandit, Kausik Chattopadhyay
Pore-forming toxins (PFTs) are typically produced as water-soluble monomers, which upon interacting with the target cells assemble into transmembrane oligomeric pores. Vibrio parahaemolyticus thermostable direct hemolysin (TDH) is an atypical PFT that exists as tetramer in solution, prior to its membrane-binding. TDH structure highlights a core beta-sandwich domain similar to those found in the eukaryotic actinoporin family of PFTs. However, TDH structure harbors an extended C-terminal region (CTR) that is not documented in the actinoporins...
October 26, 2016: Biochemical Journal
Julia M Abitbol, John J Kelly, Kevin Barr, Ashley L Schormans, Dale W Laird, Brian L Allman
Hearing loss, including noise-induced hearing loss, is highly prevalent and severely hinders an individual's quality of life, yet many of the mechanisms that cause hearing loss are unknown. The Pannexin (Panx) channel proteins, Panx1 and Panx3 are regionally expressed in many cell types along the auditory pathway and mice lacking Panx1 in specific cells of the inner ear exhibit hearing loss, suggesting a vital role for pannexins in hearing. We proposed that Panx1 and/or Panx3 null mice would exhibit severe hearing loss and increased susceptibility to noise-induced hearing loss...
October 26, 2016: Biochemical Journal
Ravi Kv Bhongir, Gopinath Kasetty, Praveen Papareddy, Matthias Morgelin, Heiko Herwald, Arne Egesten
Pseudomonas aeruginosa airway infection is common in cystic fibrosis (CF), a disease also characterized by abundant extracellular DNA (eDNA) in the airways. The eDNA is mainly derived from neutrophils accumulating in the airways and contributes to a high sputum viscosity. The altered environment in the lower airways also paves the way for chronic P. aeruginosa infection. Here, we show that mice with P. aeruginosa airway infection have increased survival and decreased bacterial load after topical treatment with DNase...
October 26, 2016: Biochemical Journal
Laura Acquasaliente, Daniele Peterle, Simone Tescari, Nicola Pozzi, Vittorio Pengo, Vincenzo De Filippis
: β2-Glycoprotein I (β2GpI) is the major autoantigen in the antiphospholipid syndrome, a thrombotic autoimmune disease. Nonetheless, the physiological role of β2GpI is still unclear. In a recent work, we have shown that β2GpI selectively inhibits the procoagulant functions of human a-thrombin (αT) (i.e. prolongs fibrin clotting time, tc, and inhibits αT-induced platelets aggregation) without affecting the unique anticoagulant activity of the protease, i.e. the proteolytic generation of the anticoagulant protein C (PC) from the PC zymogen, which interacts with αT exclusively at the protease catalytic site...
October 19, 2016: Biochemical Journal
Emilie Lukasova, Aleš Kovařík, Alena Bačíková, Martin Falk, Stanislav Kozubek
The cellular transition to senescence is associated with extensive chromatin reorganization and gene expression changes. Recent studies appeared implying an association of lamin B1 (LB1) reduction with chromatin rearrangement in human fibroblasts promoted to senescence, while the mechanisms and structural features of these relations were not yet clarified. In this work we examined the functions of LB1 and lamin B receptor (LBR) in human cancer cells. We found that both LB1 and LBR tend to deplete during cancer cells transfer to senescence by γ-irradiation...
October 19, 2016: Biochemical Journal
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