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Biochemical Journal

Marta Mikolajczak, Timothy Goodman, Mohammad K Hajihosseini
Heterozygous mutations in the gene encoding fibroblast growth factor 10 (FGF10) or its cognate receptor, FGF-receptor 2 IIIb (FGFR2-IIIb) result in two human syndromes - LADD (lacrimo-auriculo-dento-digital) and ALSG (Aplasia of lacrimal and salivary glands). To date, the partial loss-of-FGF10 function in these patients has been attributed solely to perturbed paracrine signalling functions between FGF10-producing mesenchymal cells and FGF10-responsive epithelial cells. However, the functioning of a LADD-causing G138E FGF10 mutation, which falls outside its receptor interaction interface, has remained enigmatic...
October 14, 2016: Biochemical Journal
Sukhithasri Vijayrajratnam, Anju Choorakottayil Pushkaran, Aathira Balakrishnan, Anilkumar Vasudevan, Raja Gopi Biswas, Chethampadi Gopi Mohan
Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) is an intracellular pattern recognition receptor that recognizes bacterial peptidoglycan (PG) containing meso -diaminopimelic acid ( meso DAP) and activates the innate immune system. Interestingly, few pathogenic and commensal bacteria modify their PG stem peptide by amidation of meso DAP ( meso DAPNH2). In this study, NOD1 stimulation assays were performed using bacterial PG containing meso DAP (PGDAP) and meso DAPNH2 (PGDAPNH2) to understand the differences in their biomolecular recognition mechanism...
October 14, 2016: Biochemical Journal
Ka Wai Fung, David W Wright, Jayesh Gor, Marcus J Swann, Stephen J Perkins
During the activation of complement C4 to C4b, the exposure of its thioester domain (TED) is crucial for the attachment of C4b to activator surfaces. In the C4b crystal structure, TED forms an Arg(104)-Glu(1032) salt bridge to tether its neighbouring macroglobulin (MG1) domain. Here, we examined the C4b domain structure to test whether this salt bridge affects its conformation. Dual polarisation interferometry of C4b immobilised at a sensor surface showed that the maximum thickness of C4b increased by 0.46 nm with increase in NaCl concentration from 50 mM to 175 mM NaCl...
October 13, 2016: Biochemical Journal
Alana M Thackray, Olivier Andreoletti, Raymond Bujdoso
In pursuit of a tractable bioassay to assess blood prion infectivity we have generated PrP transgenic Drosophila , which show a neurotoxic phenotype in adulthood after exposure to exogenous prions at the larval stage. Here we determined the sensitivity of ovine PrP transgenic Drosophila to ovine prion infectivity by exposure of these flies to a dilution series of scrapie-infected sheep brain homogenate. Ovine PrP transgenic Drosophila showed a significant neurotoxic response to dilutions of 10-2 through to 10-10 of the original scrapie-infected sheep brain homogenate...
October 12, 2016: Biochemical Journal
Leah D McGurn, Maryam Moazami-Goudarzi, Sean A White, Tannu Suwal, Beant Brar, Jason Q Tang, George S Espie, Matthew S Kimber
CcaA is a b-carbonic anhydrase that is a component of the carboxysomes of a subset of b-cyanobacteria. This protein, which has a characteristic C-terminal extension of unknown function, is recruited to the carboxysome via interactions with CcmM, which is itself a γ-carbonic anhydrase homolog with enzymatic activity in many, but not all cyanobacteria. We have determined the structure of CcaA from Synechocystis sp. PCC 6803 at 1.45 Å. In contrast to the dimer of dimers organization of most bacterial β-carbonic anhydrases, or the loose dimer-of-dimers-of-dimers organization found in the plant enzymes, CcaA shows a well-packed trimer-of-dimers organization...
October 11, 2016: Biochemical Journal
Shohei Iwase, Shoichiro Ono
Cyclase-associated protein (CAP) is a conserved actin-regulatory protein that functions together with actin-depolymerizing factor (ADF)/cofilin to enhance actin filament dynamics. CAP has multiple functional domains, and the function to regulate actin monomers is carried out by its C-terminal half containing a Wiskott-Aldrich Syndrome protein homology 2 (WH2) domain, a CAP and X-linked retinitis pigmentosa 2 (CARP) domain, and a dimerization motif. WH2 and CARP are implicated in binding to actin monomers and important for enhancing filament turnover...
October 11, 2016: Biochemical Journal
Adam Neal, Austin Rountree, Kelly Kernan, Brian Van Yserloo, Huiliang Zhang, Benjamin J Reed, William Osborne, Wang Wang, Ian R Sweet
A real time method to measure intracellular H2O2 would be very impactful in characterizing rapid changes that occur in physiologic and pathophysiologic states. Current methods do not provide the sensitivity, specificity and spatiotemporal resolution needed for such experiments on intact cells. We developed the use of HyPer, a genetic indicator for H2O2 that can be expressed in the cytosol (cyto-HyPer) or the mitochondria (mito-HyPer) of live cells. INS-1 cells or islets were permeabilized and the cytosolic HyPer signal was a linear function of extracellular H2O2, allowing fluorescent cyto-HyPer signals to be converted to H2O2 concentrations...
October 11, 2016: Biochemical Journal
Anna Kicinska, Bartlomiej Augustynek, Bogusz Kulawiak, Wieslawa Jarmuszkiewicz, Adam Szewczyk, Piotr Bednarczyk
Potassium channels have been found in the inner mitochondrial membrane of various cells. These channels regulate the mitochondrial membrane potential, respiration and the production of reactive oxygen species. In this study, we identified the activity of a large-conductance Ca(2+)-regulated potassium channel (mitoBKCa channel) in mitoplasts isolated from a primary human dermal fibroblast cell line. A potassium selective current was recorded with a mean conductance of 280 ± 2 pS in a symmetrical 150 mM KCl solution...
October 11, 2016: Biochemical Journal
Lei Wang, Lianzhi Li, Huaisheng Wang, Jifeng Liu
Human fibrinogen is an important coagulation factor as well as an independent predictor of coronary heart disease and stroke. Analysis of dysfibrinogens may provide useful information and help us to understanding the molecular defects in fibrin polymerization. Here the influence of oxidative stress of fibrinogen induced by H2O2 to the polymerization state of fibrin was studied. UV absorbance spectroscopy, circular dichroism, zeta-potential, dynamic light scattering, steady shear viscosity were all employed to s tudy the influence of oxidative stress to the molecular structure, the surface charges, the size and shape of fibrinogen molecules...
October 4, 2016: Biochemical Journal
Kerrie A Morrison, Aneel Akram, Ashlyn Mathews, Zoeya A Khan, Jaimin H Patel, Chumin Zhou, David John Hardy, Charles Moore-Kelly, Roshani Patel, Victor Odiba, Tim Knowles, Masood-Ul-Hassan Javed, Nikola P Chmel, Timothy R Dafforn, Alice Jane Rothnie
The use of styrene maleic acid (SMA) co-polymers to extract and purify transmembrane proteins, whilst retaining their native bilayer environment, overcomes many of the disadvantages associated with conventional detergent based procedures. This approach has huge potential for the future of membrane protein structural and functional studies. In this investigation we have systematically tested a range of commercially available SMA polymers, varying in both the ratio of styrene to maleic acid and in total size, for the ability to extract, purify and stabilise transmembrane proteins...
September 30, 2016: Biochemical Journal
Agnieszka Ruszkowska, Elzbieta Lenartowicz, Walter N Moss, Ryszard Kierzek, Elzbieta Kierzek
The influenza A virus genome is comprised of eight negative-sense viral (v)RNA segments. The seventh segment of the genome encodes two essential viral proteins and is specifically packaged alongside the other seven vRNAs. To gain insight into the possible roles of RNA structure both within and without virions, a secondary structure model of a naked (protein-free) segment 7 vRNA has been determined using chemical mapping and thermodynamic energy minimization. The proposed structure model was validated using microarray mapping, RNase H cleavage and comparative sequence analysis...
September 30, 2016: Biochemical Journal
Eriko Mishima, Yoko Sato, Kei Nanatani, Naomi Hoshi, Jong-Kook Lee, Nina Schiller, Gunnar von Heijne, Masao Sakaguchi, Nobuyuki Uozumi
Voltage-dependent potassium (Kv) channels control potassium permeability in response to shifts in the membrane potential. Voltage sensing in Kv channels is mediated by the positively charged transmembrane domain S4. The best-characterized Kv channel, KvAP, lacks the distinct hydrophilic region corresponding to the S3-S4 extracellular loop that is found in other potassium channels. Here we evaluated the topogenic properties of the transmembrane regions within the voltage-sensing domain in KvAP. S3 had low membrane insertion activity, whereas S4 possessed a unique type-I signal anchor (SA-I) function, which enabled it to insert into the membrane by itself...
September 30, 2016: Biochemical Journal
Young Ah Seo, Ruvin Kumara, Herbert A Wetli, Marianne Wessling-Resnick
Divalent Metal Transporter-1 (DMT1) mediates dietary iron uptake across the intestinal mucosa and facilitates peripheral delivery of iron released by transferrin in the endosome. Here we report that classical cannabinoids (Δ(9)-THC), non-classical cannabinoids (CP55,940), aminoalkylindoles (WIN55,212-2), and endocannabinoids (anandamide) reduce (55)Fe and (54)Mn uptake by HEK293T(DMT1) cells stably expressing the transporter. siRNA knockdown of cannabinoid receptor-2 (CB2) abrogated inhibition. CB2 is G protein-coupled receptor that negatively regulates signal transduction cascades involving serine/threonine kinases...
September 28, 2016: Biochemical Journal
Alaa Shafie, Mingzhan Xue, Guy Barker, Daniel Zehnder, Paul J Thornalley, Naila Rabbani
Glyoxalase 1 (Glo1) is a cytoplasmic enzyme with a cytoprotective function linked to metabolism of the cytotoxic side-product of glycolysis, methylglyoxal. It prevents dicarbonyl stress - the abnormal accumulation of reactive dicarbonyl metabolites increasing protein and DNA damage. Increased Glo1 expression delays ageing and suppresses carcinogenesis, insulin resistance, cardiovascular disease and vascular complications of diabetes and renal failure. Surprisingly gene trapping by the International Mouse Knockout Consortium (IMKC) to generate putative Glo1 knockout mice produced a mouse line with phenotype characterised as normal and healthy...
September 26, 2016: Biochemical Journal
Joana Fernandes Guerreiro, Alexander Muir, Subramaniam Ramachandran, Jeremy Thorner, Isabel Sá-Correia
Acetic acid-induced inhibition of yeast growth and metabolism limits the productivity of industrial fermentation processes, especially when lignocellulosic hydrolysates are used as feedstock in industrial biotechnology. Tolerance to acetic acid of food spoilage yeasts is also a problem in the preservation of acidic foods and beverages. Thus, understanding the molecular mechanisms underlying adaptation and tolerance to acetic acid stress is increasingly important in Industrial Biotechnology and Food Industry...
September 26, 2016: Biochemical Journal
Rugmani Padmanabhan Iyer, Sumin Gu, Jean X Jiang
SNAT1 is a system N/A type amino acid transporter that primarily expresses in neurons and mediates the transport of L-Glutamine. Glutamine (Gln) is an important amino acid involved in multiple cellular functions and also is a precursor for neurotransmitters, glutamate and GABA. In this study, we demonstrated that SNAT1 is an N-glycoprotein expressed in neurons. We identified three glycosylation sites at asparagine residues 251, 257 and 310 in SNAT1 protein, and that the first two are the primary sites. The biotinylation and confocal immunofluorescence analysis showed that the glycosylation-impaired mutants as well as deglycosylated SNAT1 were equally capable of expressing on the cell surface...
September 21, 2016: Biochemical Journal
Dong Keun Rhee, Jung Chae Lim, Steven C Hockman, Faiyaz Ahmad, Dong Ho Woo, Youn Wook Chung, Shiwei Liu, Allison L Hockman, Vincent C Manganiello
Oxidative stress plays a pivotal role in pathogenesis of cardiovascular diseases and diabetes, however, the roles of Protein Kinase A (PKA) and human Phosphodiesterase 3A ( hPDE3A ) remain unknown. Here we show that yeast expressing wild type (WT) hPDE3A or K13R hPDE3A (putative ubiquitinylation site mutant) exhibited resistance or sensitivity to exogenous H2O2, respectively. H2O2-stimulated ROS production was markedly increased in yeast expressing K13R hPDE3A (OxiS1), compared to yeast expressing WT hPDE3A (OxiR1)...
September 19, 2016: Biochemical Journal
Yehouda Marcus, Noam Tal, Mordechai Ronen, Raanan Carmieli, Michael Gurevitz
Ornidazole of the 5-nitroimidazole drug family is used to treat protozoan and anaerobic bacterial infections via a mechanism that involves pre-activation by reduction of the nitro group, and production of toxic derivatives and radicals. Metronidazole, another drug family member, has been suggested to affect photosynthesis by draining electrons from the electron carrier, ferredoxin, thus inhibiting NADP(+) reduction and stimulating radical and peroxide production. Here we show, however, that ornidazole inhibits photosynthesis via a different mechanism...
September 19, 2016: Biochemical Journal
Kyohei Higashi, Keita Takeda, Ann Mukuno, Yusuke Okamoto, Sayaka Masuko, Robert J Linhardt, Toshihiko Toida
Glycosaminoglycans (GAGs) including chondroitin sulfate (CS), dermatan sulfate, heparin, heparan sulfate and keratan sulfate (KS) are linear, sulfated repeating disaccharide sequences containing hexosamine and uronic acid (or galactose (Gal) in the case of KS).  Among the GAGs, CS shows structural variations, such as sulfation patterns and fucosylation, which are responsible for their physiological functions through CS interaction with CS-binding proteins.  Here, we solved the structure of KS-branched chondroitin sulfate E derived from a clam, Mactra chinensis...
September 19, 2016: Biochemical Journal
Kenneth Kao, Phillip Andrews
Pygopus 2 (Pygo2) is a chromatin effector that plays an essential role in canonical Wnt signaling associated with development and stem cell growth. Its function is to facilitate histone acetylation by recruitment of histone acetyltransferases (HATs) at active sites of β-catenin-mediated transcription. In this study, we report that Pygo2 itself, is transiently acetylated when bound to the activated TCF/β-catenin transcription complex, which correlated with β-catenin binding and Axin2 gene activation. The HATs CBP/p300, but not GCN5/PCAF targeted specific Lysine residues of the N-terminal homology domain of Pygo2 for acetylation...
September 19, 2016: Biochemical Journal
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