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93 records returned

Phosphatome profiling reveals PTPN2, PTPRJ and PTEN as potent negative regulators of PKB/Akt activation in Ras mutated cancer cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Oncogenic Ras mutations render the protein constitutively active and promote tumourigenisis via chronic stimulation of effector pathways. In A549 lung adenocarcinoma approximately 50% of the total Ras population is constitutively active yet these cells display only weak activation of the effectors: ERK1/2 and Akt. In order to identify key negative regulators of oncogenic Ras signalling we performed a phosphatome RNAi screen in A549 cells and ranked their effects on phosphorylation of Ser473 of Akt. As expected, the tumour suppressor PTEN emerged as a leading hit – knockdown elevated Akt activation to 70% of maximal ...
Source: BJ Disease - November 19, 2009 Category: Biochemistry Authors: J Omerovic, M J. Clague, I A. Prior Tags: BJ Signal Source Type: journals

MSK1 regulates the transcription of IL-1ra in response to TLR activation in macrophages.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The activity of the pro-inflammatory cytokine IL-1 is closely regulated in vivo via a variety of mechanisms, including both the control of IL-1 production and secretion as well as naturally occurring inhibitors of IL-1 function such as IL-1ra. IL-1ra is homologous to IL-1, and is able to bind but not activate the IL-1 receptor. IL-1ra can be produced by a variety of cell types, and its production is stimulated by inflammatory signals. Here we show that in macrophages the TLR mediated induction of IL-1ra from both its proximal and distal promoters involves the p38 and ERK1/2 MAPK cascades. In addition we show that mitogen a...
Source: BJ Disease - November 19, 2009 Category: Biochemistry Authors: J Darragh, O Ananieva, A Courtney, S Elcombe, J C Arthur Tags: BJ Signal Source Type: journals

CFTR fragments with the F508 deletion exert a dual allosteric control over the master kinase CK2.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Cystic Fibrosis (CF) mostly follows a single F508 deletion in CFTR (ΔF508), thereby causing premature fragmentation of the nascent protein with concomitant alterations of diverse cellular functions. We show that CK2, the most pleiotropic protein kinase, undergoes allosteric control of its different cellular forms in the presence of short CFTR peptides encompassing the F508 deletion: these ΔF508 peptides drastically inhibit the isolated catalytic subunit (α) of the kinase and yet up-regulate the holoenzyme, composed of two catalytic and two non catalytic (β) subunits. Remarkable agreement between...
Source: BJ Disease - November 19, 2009 Category: Biochemistry Authors: M A Pagano, O Marin, G Cozza, S Sarno, F Meggio, K J. Treharne, A Mehta, L A Pinna Tags: BJ Signal Source Type: journals

GDP-Mannose pyrophosphorylase is essential in the bloodstream form of Trypanosoma bruceiEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
A putative guanidine diphosphomannose pyrophophosphorylase (GDP-Man PP) gene from Trypanosoma brucei was identified in the genome and subsequently cloned, sequenced, and recombinantly expressed and shown to be a catalytically active dimer. Kinetic analysis revealed a Vmax of 0.34 μmol / min/ mg and Km’s of 67 μM and 12 μM for GTP and mannose-1-phosphate respectively. Further kinetic studies showed GDP-Man was a potent product feedback inhibitor. RNAi of the cytosolic TbGDP-Man PP showed mRNA levels were reduced to ~20% of wild type levels, causing the cells to die after 3-4 days, demonstrating ...
Source: BJ Disease - November 18, 2009 Category: Biochemistry Authors: H Denton, S Fyffe, T K Smith Tags: BJ Metabolism Source Type: journals

Microsomal prostaglandin E synthase-1 in both cancer cells and hosts contributes to tumour growth, invasion and metastasisEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Microsomal prostaglandin E synthase-1 (mPGES-1) is a stimulus-inducible enzyme that functions downstream of cyclooxygenase (COX)-2 in the prostaglandin E2 (PGE2)-biosynthetic pathway. Although COX-2-derived PGE2 is known to play a role in the development of various tumours, the involvement of mPGES-1 in carcinogenesis has not yet been fully understood. Here, we used Lewis lung carcinoma (LLC) cells with mPGES-1 knockdown or overexpression as well as mPGES-1-deficient mice to examine the roles of cancer cell- and host-associated mPGES-1 in the processes of tumorigenesis in vitro and in vivo. We found that siRNA silencing of...
Source: BJ Disease - October 20, 2009 Category: Biochemistry Authors: D Kamei, M Murakami, Y Sasaki, Y Nakatani, M Majima, Y Ishikawa, T Ishii, S Uematsu, S Akira, S Hara, I Kudo Tags: BJ Disease Source Type: journals

Comparison of the cellular and biochemical properties of Plasmodium falciparum choline and ethanolamine kinasesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The proliferation of the malaria-causing parasite Plasmodium falciparum within the erythrocyte is concomitant with massive phosphatidylcholine and phosphatidylethanolamine biosynthesis. Based on pharmacological and genetic data, de novo biosynthesis pathways of both phospholipids appear essential for parasite survival. The present study characterizes P. falciparum choline kinase (PfCK) and ethanolamine kinase (PfEK), which catalyse the first enzymatic steps of these essential metabolic pathways. Recombinant PfCK and PfEK were expressed as His-tagged fusion proteins from over-expressing E. coli strains, then purified to hom...
Source: BJ Disease - October 20, 2009 Category: Biochemistry Authors: B Alberge, L Gannoun-Zaki, C Bascunana, C Tran Van Ba, H Vial, R Cerdan Tags: BJ Metabolism Source Type: journals

Serotonin as a physiological substrate for myeloperoxidase and its superoxide dependent oxidation to cytotoxic tryptamine 4,5 dioneEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study we show that serotonin is a favoured substrate for myeloperoxidase because other physiological substrates for this enzyme, including chloride, did not affect its rate of oxidation. At low micromolar concentrations, serotonin enhanced hypochlorous acid production by both purified myeloperoxidase and neutrophils. At higher concentrations it almost completely blocked formation of hypochlorous acid. Serotonin was oxidized to a dimer by myeloperoxidase and hydrogen peroxide. It was also converted to tryptamine-4,5-dione, especially in the presence of superoxide. This toxic quinone was produced by stimulated neutro...
Source: BJ Disease - October 13, 2009 Category: Biochemistry Authors: V Ximenes, G Maghzal, R Turner, Y Kato, C C Winterbourn, A J Kettle Tags: BJ Metabolism Source Type: journals

SCG10-like protein (SCLIP) is a STAT3 interacting protein involved in maintaining epithelial morphology in MCF-7 breast cancer cells.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, we show that STAT3 binds the C-terminal tubulin associating region of SCLIP. In a search for a function of SCLIP we show that SCLIP was downregulated during OSM treatment in MCF-7 cells, which also stimulates epithelial morphology loss. SCLIP knockdown likewise triggered a loss of epithelial morphology which included reduced E-cadherin expression. We found that STAT3 was required to maintain SCLIP stability. Furthermore, inhibition of OSM-induced STAT3 activity preserved SCLIP expression and MCF-7 epithelial monolayers. Taken together, we propose that a STAT3/SCLIP interaction is required to preserved SCLIP ...
Source: BJ Disease - October 13, 2009 Category: Biochemistry Authors: D C. H. Ng, C P. Lim, B H. Lin, T Zhang, X Cao Tags: BJ Disease Source Type: journals

Role of the cysteine protease interpain A of Prevotella intermedia in breakdown and release of haem from haemoglobin.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The Gram-negative oral anaerobe Prevotella intermedia forms an iron(III) protoporphyrin IX pigment from haemoglobin. The microorganism expresses a 90 kDa cysteine protease, Interpain A (InpA), a homologue of Streptococcus pyogenes streptopain (SpeB). The role of InpA in haemoglobin breakdown and haem release was investigated. At pH 7.5, InpA mediated oxidation of oxyhaemoglobin to hydroxymethaemoglobin (in which the haem iron is oxidised to the Fe(III) state and which carries OH- as the sixth co-ordinate ligand) by limited proteolysis of globin chains as indicated by SDS-PAGE and MALDI-TOF analysis. Prolonged incubation at...
Source: BJ Disease - October 8, 2009 Category: Biochemistry Authors: D Patrick Byrne, K Wawrzonek, A Jaworska, A John Birss, J Potempa, J William Smalley Tags: BJ Structure Source Type: journals

Molecular Identification of Aspartate N-acetyltransferase and its Mutation in HypoacetylaspartiaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The brain-specific compound N-acetylaspartate (NAA) occurs almost exclusively in neurons, where its concentration reaches ≈ 20 mM. Its abundance is determined in patients by magnetic resonance spectroscopy to assess neuronal density and health. The molecular identity of the N-acetyltransferase that catalyses NAA synthesis has remained unknown, because this enzyme is membrane-bound and difficult to purify. Database searches indicated that, among the putative N-acetyltransferases (i.e., proteins homologous to known N-acetyltransferases, but with uncharacterized catalytic activity) encoded by the human and mouse genome...
Source: BJ Disease - October 5, 2009 Category: Biochemistry Authors: E Wiame, D Tyteca, N Pierrot, F Collard, M Amyere, G Noël, J Desmedt, M Nassogne, M Vikkula, J Octave, M Vincent, P J Courtoy, E Boltshauser, E Van Schaftingen Tags: BJ Metabolism Source Type: journals

Substrate specificity and inhibitors of LRRK2, a protein kinase mutated in Parkinson{'}s diseaseEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The Leucine Rich Repeat Protein Kinase-2 (LRRK2) is mutated in a significant number of Parkinson’s disease patients, but little is known about its regulation and function. A common mutation changing Gly2019 to Ser enhances catalytic activity, suggesting small molecule inhibitors might have utility in treating Parkinson’s Disease. We utilised various approaches to explore the substrate specificity requirements of LRRK2 and elaborated a peptide substrate termed Nictide, that had 20-fold lower Km and nearly 2-fold higher Vmax than the widely deployed LRRKtide substrate. We demonstrate that LRRK2 has marked prefe...
Source: BJ Disease - September 8, 2009 Category: Biochemistry Authors: R Jeremy Nichols, N Dzamko, J E. Hutti, L C. Cantley, M Deak, J Moran, P Bamborough, A D. Reith, D R. Alessi Tags: BJ Signal Source Type: journals

Absence of iron-regulatory protein Hfe results in hyperproliferation of retinal pigment epithelium: role of cystine/glutamate exchangerEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Hemochromatosis is an iron overload disorder with age-dependent oxidative stress and dysfunction in a variety of tissues. Mutations in HFE are responsible for most cases with hemochromatosis. We recently demonstrated that Hfe is expressed exclusively in the basal membrane of retinal pigment epithelium (RPE). Here we used Hfe-/- mice to examine ferritin levels (an indirect readout for iron levels) and morphological changes in retina. We found increased ferritin accumulation in retina in 18-month-old but not in 2-month-old mice with considerable morphological damage compared to age-matched controls. The retinal phenotype inc...
Source: BJ Disease - August 27, 2009 Category: Biochemistry Authors: J P. Gnanaprakasam, M Thangaraju, K Liu, Y Ha, P M. Martin, S B. Smith, V Ganapathy Tags: BJ Disease Source Type: journals

MAC inhibitors suppress mitochondrial apoptosisEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
MAC (Mitochondrial Apoptosis-induced Channel) forms in the mitochondrial outer membrane and unleashes cytochrome c, which then orchestrates the execution of the cell. MAC opening is the commitment step of intrinsic apoptosis. Hence, closure of MAC may prevent apoptosis. Compounds that blocked release of fluorescein from liposomes by recombinant Bax were tested for their ability to directly close MAC and suppress apoptosis in FL5.12 cells. Low doses of these compounds (IC50 ranged from 19 to 966 nM) irreversibly closed MAC. These compounds also blocked cytochrome c release and halted the onset of apoptotic markers normally ...
Source: BJ Disease - August 19, 2009 Category: Biochemistry Authors: P M. Peixoto, S Ryu, A Bombrun, B Antonsson, K W. Kinnally Tags: BJ Cell Source Type: journals

Biochemical consequences of sedlin mutations that cause spondyloepiphyseal dysplasia tardaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Spondyloepiphyseal dysplasia tarda (SEDT) is a late-onset X-linked recessive skeletal dysplasia caused by mutations in the gene SEDL coding for sedlin. Here, we investigate four missense mutations observed in SEDT and compare biochemical and cellular characteristics relative to the wild-type protein to address mechanism of disease and to gain insight into the function of the sedlin protein. In situ hybridization and immunohistochemical experiments in mouse growth plates revealed sedlin to be predominantly expressed in proliferating and hypertrophic chondrocytes. Cell culture studies showed the wild-type protein localized p...
Source: BJ Disease - August 2, 2009 Category: Biochemistry Authors: M Y. Choi, C C.Y. Chan, D Chan, K D.K. Luk, K S.E. Cheah, J A. Tanner Tags: BJ Disease Source Type: journals

Pyruvate dehydrogenase kinase isoenzyme 4 (PDHK4) deficiency attenuates the long-term negative effects of a high-saturated fat dietEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The hypothesis that PDHK4 has potential as a target for the treatment of type 2 diabetes was tested by feeding wild type and PDHK4 knockout mice a high saturated fat diet that induces hyperglycemia, hyperinsulinemia, glucose intolerance, hepatic steatosis, and obesity. Previous studies have shown that PDHK4 deficiency lowers blood glucose by limiting the supply of three carbon gluconeogenic substrates to the liver. There is concern, however, that the increase in glucose oxidation caused by less inhibition of the pyruvate dehydrogenase complex by phosphorylation will inhibit fatty acid oxidation, promote ectopic fat accumul...
Source: BJ Disease - July 22, 2009 Category: Biochemistry Authors: B Hwang, N Jeoung, R A Harris Tags: BJ Metabolism Source Type: journals

Promoting assembly and bundling of FtsZ as a strategy to inhibit bacterial cell division: a new approach for developing novel antibacterial drugsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
FtsZ plays an essential role in bacterial cell division. We have used the assembly of FtsZ as a screen to find antibacterial agents with a novel mechanism of action. The effects of 81 compounds of 29 different structural scaffolds on FtsZ assembly in vitro were examined using a sedimentation assay. Out of these 81 compounds, 3-{5-[4-Oxo-2-thioxo-3-(3-trifluoromethyl-phenyl)-thiazolidin-5-ylidenemethyl]-furan-2-yl}-benzoic acid (OTBA) was found to promote FtsZ assembly in vitro. OTBA increased the assembly of FtsZ, caused bundling of FtsZ protofilaments, prevented dilution-induced disassembly of FtsZ protofilaments and decr...
Source: BJ Disease - July 6, 2009 Category: Biochemistry Authors: T K Beuria, P Singh, A Surolia, D Panda Tags: BJ Cell Source Type: journals

Endogenous spartin (SPG20) is recruited to endosomes and lipid droplets and interacts with the ubiquitin E3 ligases AIP4 and AIP5.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The hereditary spastic paraplegias (HSPs) are genetic conditions exhibiting distal degeneration of the longest axons of the corticospinal tract, resulting in spastic paralysis of the legs. The gene encoding spartin is mutated in Troyer syndrome, an HSP in which paralysis is accompanied by additional clinical features. There has been controversy over the subcellular distribution of spartin. We show here that at steady state endogenous spartin exists in a cytosolic pool that can be recruited to endosomes and to lipid droplets. Cytosolic endogenous spartin is mono-ubiquitinated and we demonstrate that it interacts via a PPXY ...
Source: BJ Disease - July 5, 2009 Category: Biochemistry Authors: T L Edwards, V E Clowes, H T H Tsang, J W Connell, C M Sanderson, J Paul Luzio, E Reid Tags: BJ Cell Source Type: journals

Prostaglandin EP2 and EP4 receptors modulate CCL2 (MCP-1) expression in response to LPS-induced renal glomerular inflammationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The pro-inflammatory chemokine CCL2 (MCP-1) is upregulated in the glomerular compartment during the early phase of LPS-induced nephritis. This upregulation also appears in cultured mesangial cells (MC) and is more pronounced in MC lacking the prostaglandin EP2 receptor or in MC treated with a prostaglandin EP4 receptor antagonist. To examine a possible feedback of EP receptor stimulation on CCL2 expression, we chose an in vitro model in MC with down regulated EP receptor expression. By selective overexpression of EP receptors in these cells, their effects on LPS-induced CCL2 expression were examined. Cells were stimulated...
Source: BJ Disease - July 1, 2009 Category: Biochemistry Authors: G Zahner, M Schaper, U Panzer, M Andreas Kluger, R A.K. Stahl, F Thaiss, A Schneider Tags: BJ Cell Source Type: journals

Chemokine degradation by the Group A streptococcal serine proteinase ScpC can be reconstituted in vitro and requires two separate domainsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Streptococcus pyogenes is one of the most common human pathogens and possesses diverse mechanisms to evade the human immune defence. One example of its immune evasion is the degradation of the chemokine IL-8 by ScpC, a serine proteinase that prevents the recruitment of neutrophils to an infection site. By applying the ANTIGENome technology and using human serum antibodies we identified Spy0416, annotated as ScpC, as a prominent antigen that induces protective immune responses in animals. We demonstrate here for the first time that the recombinant form of Spy0416 is capable of IL-8 degradation in vitro in a concentration an...
Source: BJ Disease - June 23, 2009 Category: Biochemistry Authors: A Fritzer, B Noiges, D Schweiger, A Rek, A J. Kungl, A von Gabain, E Nagy, A L. Meinke Tags: BJ Disease Source Type: journals

Ephrin-independent regulation of cell substrate adhesion by the EphB4 receptorEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Receptor tyrosine kinases of the Eph family become tyrosine phosphorylated and initiate signaling events upon binding of their ligands, the ephrins. Eph receptors such as EphA2 and EphB4 are highly expressed but poorly tyrosine phosphorylated in many types of cancer cells, suggesting a limited interaction with ephrin ligands. Nevertheless, decreasing the expression of these receptors affects the malignant properties of cancer cells, suggesting that Eph receptors may influence cancer cells independently of ephrin stimulation. Ligand-independent activities of Eph receptors in cancer, however, have not been demonstrated. By u...
Source: BJ Disease - June 23, 2009 Category: Biochemistry Authors: N K. Noren, N Yang, M Silldorff, R Mutyala, E B. Pasquale Tags: BJ Cell Source Type: journals

Recombinant production of eight human cytosolic aminotransferases and assessment of their potential involvement in glyoxylate metabolismEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Primary hyperoxaluria type 1 (PH1) is a severe inborn disorder of glyoxylate metabolism, caused by a functional deficiency of the peroxisomal enzyme alanine:glyoxylate aminotransferase (AGXT), which converts glyoxylate to glycine using L-alanine as the amino-group donor. Even though pre-genomic studies indicate that other human transaminases can convert glyoxylate to glycine, in PH1 patients these enzymes are apparently unable to compensate for the lack of AGXT - perhaps due to their limited levels of expression, or to their localization in an inappropriate cell compartment, or to the scarcity of the required amino-group d...
Source: BJ Disease - June 22, 2009 Category: Biochemistry Authors: S Donini, M Ferrari, C Fedeli, M Faini, I Lamberto, A Marletta, L Mellini, M Panini, R Percudani, L Pollegioni, L Caldinelli, S Petrucco, A Peracchi Tags: BJ Disease Source Type: journals

Exposure of HL-60 human leukemic cells to 4-hydroxynonenal promotes the formation of adduct(s) with {alpha}-enolase devoid of plasminogen binding activityEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
4-Hydroxynonenal (HNE), the major product of lipoperoxidation, easily reacts with proteins through adduct formation between its three main functional groups and lysyl, histidyl, and cysteinyl residues of proteins. HNE is considered to be an ultimate mediator of toxic effects elicited by oxidative stress. It can be detected in several patho-physiological conditions, in which it affects cellular processes by addition to functional proteins. We demonstrated by mass spectrometry and confirmed by immunoblotting experiments the formation of HNE-α-enolase adduct(s) in HL-60 human leukemic cells. α-Enolase is a multi...
Source: BJ Disease - June 9, 2009 Category: Biochemistry Authors: F Gentile, S Pizzimenti, A Arcaro, P Pettazzoni, R Minelli, D D'Angelo, G Mamone, P Ferranti, C Toaldo, G Cetrangolo, S Formisano, M Umberto Dianzani, K Uchida, C Dianzani, G Barrera Tags: BJ Disease Source Type: journals

Hypothiocyanous acid reactivity with low-molecular-mass and protein thiols: Absolute rate constants and assessment of biological relevanceEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study shows for the first time that the reactions of HOSCN with low-molecular-mass thiol residues occur with rate constants in the range 7.3 × 103 M–1 s–1 (for N-acetyl-cysteine) at pH 7.4 to 7.7 × 106 M–1 s–1 (for 5-thio-2-nitrobenzoic acid at pH 6.0). An inverse relationship between the rate of reaction and the pKa of the thiol group was observed. The rates of reaction of HOSCN with thiol-containing proteins were also investigated for four proteins (creatine kinase, bovine serum albumin, β−lactoglobulin and crystallins). The values obtained for Cys residues on ...
Source: BJ Disease - June 3, 2009 Category: Biochemistry Authors: O Skaff, D I. Pattison, M J. Davies Tags: BJ Disease Source Type: journals

Deletion of the mouse Slc30a8 gene encoding zinc transporter-8 results in impaired insulin secretionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The Slc30a8 gene encodes the islet-specific zinc transporter ZnT-8, which provides zinc for insulin-hexamer formation. Polymorphic variants in amino acid 325 of human ZnT-8 are associated with altered susceptibility to type 2 diabetes and ZnT-8 autoantibody epitope specificity changes in type 1 diabetes. To assess the physiological importance of ZnT-8, mice carrying a Slc30a8 exon 3 deletion were analyzed histologically and phenotyped for energy metabolism and pancreatic hormone secretion. No gross anatomical or behavioral changes or differences in body weight were observed between wild type and ZnT-8 -/- mice and ZnT-8 -/...
Source: BJ Disease - May 18, 2009 Category: Biochemistry Authors: L D Pound, S Sarkar, R KP Benninger, Y Wang, A Suwanichkul, M K Shadoan, R L Printz, J K Oeser, C E Lee, D W Piston, O P McGuinness, J C Hutton, D R Powell, R Mark O'Brien Tags: BJ BJ Disease Source Type: journals

Dynasore inhibits removal of wild-type and {Delta}F508 CFTR from the plasma membraneEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Dynasore, a small molecule inhibitor of dynamin, was used to probe the role of dynamin in the endocytosis of wild-type and mutant CFTR. Internalization of both wild-type and “temperature-corrected” ΔF508 CFTR was markedly inhibited by short exposure to dynasore, implicating dynamin as a key element in the endocytic internalization of both wild-type and mutant CFTR. The inhibitory effect of dynasore was readily reversible upon washout of dynasore from the growth media. Corr-4, a pharmacological corrector of ΔF508 CFTR biosynthesis, caused a marked increase in the cell surface expression of mutant...
Source: BJ Disease - May 14, 2009 Category: Biochemistry Authors: A Young, M Gentzsch, C Y. Abban, Y Jia, P I. Meneses, R J. Bridges, N A. Bradbury Tags: BJ BJ Disease Source Type: journals

NADPH oxidase-mediated reactive oxygen species alter astrocyte membrane molecular order via phospholipase A2Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We examined ROS production in astrocytes and downstream effects leading to changes in signaling cascade, morphology, and membrane dynamics using menadione, a redox active compound capable of inducing intracellular ROS. NADPH oxidase mediated menadione-induced ROS production, which then stimulated phosphorylation of p38 mitogen-activated protein kinase (MAPK) and extracellular-signal regulated kinases (ERK1/2), and increased actin polymerization and cytoskeletal protrusions. We also showed that astrocyte plasma membranes became more molecularly-ordered under oxidative stress, which was abrogated by down regulating cytosolic...
Source: BJ Disease - April 24, 2009 Category: Biochemistry Authors: D Zhu, C Hu, W Sheng, K S Tan, M A Haidekker, A Y Sun, G Y Sun, J C-M Lee Tags: BJ Cell Source Type: journals

Identification of pancreatic stellate cell population with properties of progenitor cells: new role for stellate cells in pancreasEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Numerous studies conducted in a diversity of adult tissues have reported that certain stem cells are characterized by the expression of a protein known as the ABCG2 transporter. In the adult pancreas, various multipotent progenitors have been proposed, however the ABCG2 marker has only been detected in the so-called Side Population. In the present study we aim to identify new ABCG2+ pancreatic cell populations and to explore whether they exhibit properties of progenitor cells. We isolated and expanded mitoxantrone-resistant cells from pancreata of lactating rats by drug selection. These cells were characterized an...
Source: BJ Disease - April 20, 2009 Category: Biochemistry Authors: E Mato, M Lucas, J Pétriz, R Gomis, A Novials Tags: BJ Cell Source Type: journals

Two pathways for cysteine biosynthesis in Leishmania majorEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Genome mining and biochemical analyses have shown that L. major possesses two pathways for cysteine synthesis - the de novo biosynthesis pathway comprising serine acetyltransferase (SAT) and cysteine synthase (CS) and the reverse transsulfuration (RTS) pathway comprising cystathionine β-synthase (CBS) and cystathionine gamma-lyase (CGL). The L. major CS (LmjCS) is similar to the type A CSs of bacteria and catalyses the synthesis of cysteine using O-acetyserine and sulfide with Kms of 17.5 and 0.13 mM, respectively. LmjCS can use sulfide provided by the action of mercaptopyruvate sulfurtransferase (MST) on 3-mercapto...
Source: BJ Disease - March 18, 2009 Category: Biochemistry Authors: R A M Williams, G D Westrop, G H Coombs Tags: BJ Cell Source Type: journals

Hydrogen sulfide destroys lipid hydroperoxides in oxidized LDLEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Lipid hydroperoxides (LOOHs) in oxidized LDL (oxLDL) are potentially atherogenic compounds. Recently, H2S was identified as the third endogenous gasotransmitter in the vasculature. Hydrogen peroxide is known to be destroyed by H2S. Assuming that H2S may also react with LOOHs, the results show that H2S can destroy LOOHs in oxLDL. LOOH-enriched LDL after H2S-pretreatment was not further oxidized by endothelial cells and macrophages and the ability of oxLDL to induce HO-1 in endothelial cells was abolished by H2S pretreatment. HPLC analysis showed that 9-HPODE (9-hydroperoxy-octadecadienoic acid), a compound found in oxLDL, w...
Source: BJ Disease - March 5, 2009 Category: Biochemistry Authors: M K. Muellner, S M. Schreier, H Laggner, M Hermann, H Esterbauer, M Exner, B M.K. Gmeiner, S Kapiotis Tags: BJ Disease Source Type: journals

Identification of small molecule inhibitors of the JIP and JNK interactionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The c-Jun N-terminal kinase 1 (JNK1) plays a crucial role in the regulation of obesity-induced insulin resistance and is implicated in the pathology of type II diabetes. Its partner, JNK interacting protein 1 (JIP1), serves a scaffolding function facilitating JNK1 activation by mitogen-activated protein (MAP) kinase kinase 4 (MKK4) and MAP kinase kinase 7 (MKK7). For example, reduced insulin resistance and JNK activation are observed in JIP1 deficient mice. Based on the in vivo efficacy of a cell-permeable JIP peptide, the JIP/JNK interaction appears to be a potential target for JNK inhibition. The goal of this study was t...
Source: BJ Disease - February 25, 2009 Category: Biochemistry Authors: T Chen, N Kablaoui, J Little, S Timofeevski, W R Tschantz, P Chen, J Feng, M Charlton, R Stanton, P Bauer Tags: BJ Disease Source Type: journals

Biochemical evidence of the interactions of membrane type-1 matrix metalloproteinase (MT1-MMP) with ADP/ATP nucleoside transporter (ANT): potential implications linking proteolysis with the energy metabolism in cancer cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Invasion-promoting membrane type-1 matrix metalloproteinase (MT1-MMP) is a key element in cell migration processes. To identify the proteins, which interact and, therefore, co-precipitate with this proteinase from cancer cells, we used the proteolytically potent wild-type (WT), the catalytically inert E240A (E240A) and the C-end truncated (tailless; ΔCT) MT1-MMP-FLAG constructs as baits. The identity of the pulled-down proteins was determined by LC-MS/MS and then confirmed with specific antibodies. We determined that in breast carcinoma MCF-7 cells ADP/ATP nucleoside transporter (ANT) efficiently interacted with the...
Source: BJ Disease - February 20, 2009 Category: Biochemistry Authors: I A. Radichev, A G. Remacle, N Sounni, S A. Shiryaev, D V. Rozanov, W Zhu, N V. Golubkova, T I. Postnova, V S. Golubkov, A Y. Strongin Tags: BJ Cell Source Type: journals

The regulation of the intestinal mucin MUC2 expression by short chain fatty acids: implications for epithelial protectionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In conclusion, 1 mM of butyrate and 1-15 mM of propionate increase MUC2 expression. The effects of butyrate on MUC2 mRNA are mediated via AP-1 and acetylation/methylation of histones at the MUC2 promoter. (Source: BJ Disease)
Source: BJ Disease - February 19, 2009 Category: Biochemistry Authors: N Burger-van Paassen, A Vincent, P J Puiman, M van der Sluis, J Bouma, G Boehm, J B van Goudoever, I Van Seuningen, I B Renes Tags: BJ Gene Source Type: journals

Lipid phosphate phosphohydrolase type 1 (LPP1) degrades extracellular lysophosphatidic acid in vivoEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We report here the development of a mouse wherein the LPP1 gene (Ppap2a) was disrupted. The homozygous mice, which are phenotypically unremarkable, generally lack LPP1 mRNA and multiple tissues exhibit a substantial (35 – 95%) reduction in LPA phosphatase activity. Compared to wild type littermates, Ppap2atr/tr animals have increased levels of plasma LPA and LPA injected intravenously is metabolized at a four-fold slower rate. Our results demonstrate that LPA is rapidly metabolized in the bloodstream and that LPP1 is an important determinant of this turnover. These results indicate that LPP1 is a catabolic enzyme fo...
Source: BJ Disease - February 12, 2009 Category: Biochemistry Authors: J L. Tomsig, A H. Snyder, E V. Berdyshev, A Skobeleva, C Mataya, V Natarajan, D N. Brindley, K R. Lynch Tags: BJ Signal Source Type: journals

Post-translational import of protein into the endoplasmic reticulum of a trypanosome: an in vitro system for discovery of anti-trypanosomal chemical entitiesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Human African trypanosomiasis (HAT), caused by the protozoan parasite Trypanosoma brucei, is an emerging disease for which new drugs are needed. Expression of plasma membrane proteins, (e.g., variant surface glycoprotein (VSG)), is crucial for establishment and maintenance of an infection by T. brucei. Transport of a majority of proteins to the plasma membrane involves their translocation into the endoplasmic reticulum (ER). Thus, inhibition of protein import into the ER of T. brucei would be a logical target for discovery of lead compounds against trypanosomes. We have developed a trypanosome microsomal system (TbRM) that...
Source: BJ Disease - February 5, 2009 Category: Biochemistry Authors: B Patham, J Duffy, A Lane, R C. Davis, P Wipf, S W. Fewell, J L. Brodsky, K Mensa-Wilmot Tags: BJ Cell Source Type: journals

Expression of the iron-regulatory protein hemojuvelin in retina and its regulation during cytomegalovirus infectionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Hemochromatosis is a genetic disorder of iron overload resulting from loss-of-function mutations in genes coding for the iron-regulatory proteins HFE, transferrin receptor 2, ferroportin, hepcidin, and hemojuvelin (HJV). Expression of the first four genes in retina has been established. Here we report on the expression of HJV. Since infection of retina with cytomegalovirus (CMV) causes blindness, we also investigated the expression of HJV and other iron-regulatory proteins in retina during CMV infection. Hjv mRNA was expressed in RPE/eyecup and neural retina in mouse. In situ hybridization and immunohistochemistry confirme...
Source: BJ Disease - February 4, 2009 Category: Biochemistry Authors: J P. Gnana-Prakasam, M Zhang, P M. Martin, S S. Atherton, S B. Smith, V Ganapathy Tags: BJ Cell Source Type: journals

Structural and functional characterization of SplA, an exclusively specific protease of Staphylococcus aureusEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Staphylococcus aureus is a dangerous human pathogen which antibiotic resistance is steadily increasing and no efficient vaccine is as yet available. This serious threat drives extensive studies on staphylococcal physiology and pathogenicity pathways, especially virulence factors. Serine protease like proteins (Spl) encoded by an operon containing up to six genes are a good example of poorly characterized secreted proteins likely involved in virulence. Here we describe an efficient heterologous expression system for SplA and detailed biochemical and structural characterization of the recombinant SplA protease. The enzyme sh...
Source: BJ Disease - January 28, 2009 Category: Biochemistry Authors: J Stec-Niemczyk, K Pustelny, M Kisielewska, M Bista, K T. Boulware, H R. Stennicke, I B. Thogersen, P S. Daugherty, J J. Enghild, G M. Popowicz, A Dubin, J Potempa, G Dubin Tags: BJ Structure Source Type: journals

The soluble metal-binding domain of copper transporter ATP7B binds and detoxifies cisplatinEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Wilson disease ATPase (ATP7B) has been implicated in the resistance of cancer cells to cisplatin. Using a simple in vivo assay in bacterial culture, we demonstrate that ATP7B can confer resistance to cisplatin by sequestering the drug in its N-terminal metal-binding domain without active drug extrusion from the cell. Expression of a protein fragment containing four N-terminal metal binding repeats of ATP7B (MBR1-4) protects cells from the toxic effects of cisplatin. One MBR1-4 molecule binds up to three cisplatin molecules at the copper-binding sites in the metal-binding repeats. These findings suggest that suppressing enz...
Source: BJ Disease - January 27, 2009 Category: Biochemistry Authors: N V Dolgova, D Olson, S Lutsenko, O Y Dmitriev Tags: BJ Disease Source Type: journals

Retinoids synergize with insulin to induce hepatic Gck ExpressionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Hepatic glucokinase (GK) plays a key role in maintaining glucose homeostasis. Many stimuli regulate GK activity by controlling its gene transcription. We hypothesized that endogenous lipophilic molecules modulate hepatic Gck expression. Lipophilic molecules were extracted from rat livers, saponified and re-constituted as a lipophilic extract (LE). LE synergized with insulin to induce primary hepatocyte, but not beta-cell, Gck expression in a sterol regulatory element-binding protein 1c (SREBP-1c) independent manner. The dramatic induction of Gck mRNA resulted in a significant increase of GK activity. Subsequently, the acti...
Source: BJ Disease - January 27, 2009 Category: Biochemistry Authors: G Chen, Y Zhang, D Lu, N Li, A Catharine Ross Tags: BJ Disease Source Type: journals

Analysis and characterization of dimerization inhibition of a multi-drug resistant Human Immunodeficiency Type 1 protease using a novel size exclusion chromatographic approachEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Active site inhibitors of human immunodeficiency type 1 (HIV-1) protease (PR) block viral replication by preventing viral maturation. However, HIV-1 often develops resistance to active site inhibitors through multiple mutations in PR and therefore recent efforts have focused on inhibiting PR dimerization as an alternative approach. Dimerization inhibitors have been identified using kinetic analysis but additional characterization of the effect of these inhibitors on PR by physical methods has been difficult. Here, we identified a multi-drug resistant HIV-1 PR (PRMDR) that was highly resistant to autoproteolysis. Using this...
Source: BJ Disease - January 16, 2009 Category: Biochemistry Authors: D A Davis, I R Tebbs, S I. Daniels, S J Stahl, J D Kaufman, P Wingfield, M J Bowman, J Chmielewski, R Yarchoan Tags: BJ Structure Source Type: journals

A low resolution study of the ultrastructure of Fragile X related proteinsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Fragile X related proteins form a family implicated in RNA metabolism. Their sequence is composed of conserved N-terminal and central regions which contain Tudor and KH domains and of a divergent C-terminus with motifs rich in Arg and Gly. The most interesting member of the family is probably FMRP, since absence or mutation of this protein in human causes fragile X syndrome, the most common cause of inherited mental retardation. Understanding the structural properties of FMRP is essential for correlating it to its functions. The structures of isolated domains of FMRP have been reported, but nothing is yet known about the s...
Source: BJ Disease - January 14, 2009 Category: Biochemistry Authors: L Sjekloća, P V Konarev, J Eccleston, I A Taylor, D I Svergun, A Pastore Tags: BJ Structure Source Type: journals

Chondroitin 4-O-sulfotransferase-1 is required for somitic muscle development and motor axon guidance in zebrafishEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Chondroitin sulfate (CS) has been implicated in a variety of biological processes during development. Its biological functions are closely associated with characteristic sulfated structures. Here, we report the characterization of a zebrafish counterpart of chondroitin 4-O-sulfotransferase-1 (C4ST-1) and its functional importance in embryogenesis. Recombinant C4ST-1 showed a substrate preference for chondroitin and catalyzed the 4-O-sulfation of GalNAc residues, a highly frequent modification of CS in the embryos of zebrafish as well as other vertebrates. Whole-mount in situ hybridization revealed that C4ST-1 showed a dist...
Source: BJ Disease - January 6, 2009 Category: Biochemistry Authors: S Mizumoto, T Mikami, D Yasunaga, N Kobayashi, H Yamauchi, A Miyake, N Itoh, H Kitagawa, K Sugahara Tags: BJ Structure Source Type: journals

Peroxiredoxin 4 knockout results in elevated spermatogenic cell death via oxidative stressEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Peroxiredoxin (Prx) is a multifunctional redox protein with thioredoxin-dependent peroxidase activity. Prx4 is present as a secretory protein in most tissues, whereas in sexually mature testes it is anchored in the endoplasmic reticulum (ER) membrane of spermatogenic cells via an uncleaved N-terminal hydrophobic peptide. We generated a Prx4-knockout mouse to investigate the function of Prx4 in vivo. Prx4−/y mice lacking Prx4 expression in all cells were obtained by mating Prx4flox/+ female mice with Cre-transgenic male mice that ubiquitously expressed Cre recombinase. The resulting Prx4−/y male mice were fert...
Source: BJ Disease - December 23, 2008 Category: Biochemistry Authors: Y Iuchi, F Okada, S Tsunoda, N Kibe, N Shirasawa, M Ikawa, M Okabe, Y Ikeda, J Fujii Tags: BJ Energy Source Type: journals

Membrane-targeted synergistic activity of docosahexaenoic acid and lysozyme against P. aeruginosaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Antimicrobial polypeptides including lysozymes (Ly) have membrane perturbing activity and are well documented effector molecules of innate immunity. In cystic fibrosis, a hereditary disease with frequent lung infection with Pseudomonas aeruginosa, the free fatty acid docosahexaenoic acid (DA), but not oleic acid (OA), is decreased and DA supplementation has been shown to improve the clinical condition in these patients. We hypothesized that DA may alone, or in conjunction with Ly, exert antibacterial action against P. aeruginosa. We found that DA and Ly synergistically inhibit the metabolic activity of P. aeruginosa, in co...
Source: BJ Disease - December 23, 2008 Category: Biochemistry Authors: J G. Martinez, M Waldon, Q Huang, S Alvarez, A Oren, N Sandoval, M Du, F Zhou, A Zenz, K Lohner, R Desharnais, E Porter Tags: BJ Structure Source Type: journals

Lysine-specific gingipain promotes lipopolysaccharide- and active vitamin D3-induced osteoclast differentiation by degrading osteoprotegerinEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, we examined the effects of gingipains on osteoclast differentiation. In co-cultures of mouse bone marrow cells and osteoblasts, formation of multinucleated osteoclasts induced by 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3] was augmented by Kgp but not by RgpB. A physiological concentration (0.1 nM) of 1α,25(OH)2D3 induced the osteoclast formation in the presence of 100 nM Kgp to the extent comparable to that induced by 10 nM 1α,25(OH)2D3. Kgp also enhanced osteoclastogenesis induced by various microbial components including lipopolysaccharide. Combined use of Kgp and 1α,25(OH)...
Source: BJ Disease - December 22, 2008 Category: Biochemistry Authors: R Yasuhara, Y Miyamoto, M Takami, T Imamura, J Potempa, K Yoshimura, R Kamijo Tags: BJ Cell Source Type: journals

Phosphorylation of EWS and EWS-Fli1 in response to DNA damageEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In Ewing’s sarcomas, chromosomal translocations cause the N-terminal domain of the Ewing’s sarcoma protein (EWS) to fuse with the DNA-binding domains of the Ets family of transcription factors. Here we show that EWS and EWS-Fli1, the fusion most frequently found in Ewing’s sarcomas, become phosphorylated at Thr79 in response to either mitogens or DNA damaging agents. The much weaker mitogen-induced phosphorylation of EWS is catalysed by the MAP kinases ERK1 and ERK2, while the much stronger phosphorylation of EWS induced by the DNA alkylating agent methyl methanesulphonate (MMS) can be catalysed by JNK...
Source: BJ Disease - December 16, 2008 Category: Biochemistry Authors: I V Klevernic, S Morton, R J Davis, P Cohen Tags: BJ Signal Source Type: journals

Copper proteins and ferroxidases in human plasma and that of wild type and ceruloplasmin knockout miceEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In the blood plasma of humans and rats, ceruloplasmin is the major Cu-binding protein and ferroxidase, accounting for 70% of the copper, the rest binding primarily to albumin and a macroglobulin. Systematic studies with fresh plasma were carried out to compare what occurs in the mouse. C57BL6 mice had half as much copper and p-phenylene diamine (pPD) oxidase activity as humans and rats; 20-40% as much ferroxidase activity as humans (by three different assays); and less inhibition by azide. Plasma of ceruloplasmin knockout mice had no pPD oxidase activity, but retained >50% ferroxidase activity (not as affected by azide). M...
Source: BJ Disease - December 16, 2008 Category: Biochemistry Authors: L W Gray, T Z Kidane, A Nguyen, S Akagi, K Petrasek, Y Chu, A Cabrera, K Kantardjieff, A Z Mason, M C Linder Tags: BJ Disease Source Type: journals

Antizyme-3 inhibits polyamine uptake and ornithine decarboxylase (ODC) activity, but does not stimulate ODC degradationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Antizymes are small polyamine-induced proteins that function as feedback regulators of cellular polyamine homeostasis. They bind to transient ornithine decarboxylase (ODC) monomeric subunits, resulting in inhibition of ODC activity and targeting ODC to ubiquitin-independent proteasomal degradation. Antizyme 3 (Az3) is a mammalian antizyme isoform expressed exclusively in testicular germ cells and therefore considered as a potential regulator of polyamines during spermatogenesis. We show here that unlike antizyme 1 (Az1) and antizyme 2 (Az2), which efficiently inhibit ODC activity and stimulate its proteasomal degradation, ...
Source: BJ Disease - December 15, 2008 Category: Biochemistry Authors: Z Snapir, A Keren-Paz, Z Bercovich, C Kahana Tags: BJ Disease Source Type: journals

The C-terminus of GLUT4 targets the transporter to the perinuclear compartment but not to the insulin-responsive vesiclesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Postprandial blood glucose clearance is mediated by glucose transporter isoform 4 (Glut4) which is translocated from an intracellular storage pool to the plasma membrane in response to insulin. The nature of the intracellular storage pool of Glut4 is not well understood. Immunofluorescence staining shows that, under basal conditions, the major population of Glut4 resides in the perinuclear compartment. At the same time, biochemical fractionation reveals that Glut4 is localized in insulin-responsive vesicles, or IRVs. The relationship between the perinuclear Glut4 compartment and the IRVs is not known. Here, we have exchang...
Source: BJ Disease - December 15, 2008 Category: Biochemistry Authors: L V. Li, K Bakirtzi, R T. Watson, J E. Pessin, K Kandror Tags: BJ Cell Source Type: journals

Trafficking of immature {Delta}F508-CFTR to the plasma membrane and its detection by biotinylationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Recent studies suggest that immature, core-glycosylated ΔF508-CFTR (the predominant mutant form of the Cystic Fibrosis Transmembrane conductance Regulator) can reach the plasma membrane under some conditions. We investigated this possibility since it has implications for understanding how therapeutics rescue the trafficking of mutant CFTR and perhaps other misfolded proteins. Core-glycosylated CFTR was labelled and pulled down on streptavidin beads after exposure to sulfo-NHS-SS-biotin (Mr = 606.7) however intracellular proteins were also detected in the precipitates. When the R domain of CFTR was expressed in the c...
Source: BJ Disease - December 4, 2008 Category: Biochemistry Authors: Y Luo, K McDonald, J W Hanrahan Tags: BJ Cell Source Type: journals

Insulin resistance in polycystic ovary syndrome is associated with defective regulation of ERK1/2 by insulin in skeletal muscle in vivoEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study compares the major insulin signalling pathways in skeletal muscle isolated from PCOS and controls. We measured whole body insulin sensitivity and insulin signalling in skeletal muscle biopsies taken before and after acute exposure to hyperinsulinaemia in nine women diagnosed with PCOS and seven controls. We examined the expression, basal activity and response to in vivo insulin stimulation of three signalling molecules within these human muscle samples, namely, insulin receptor substrate-1 (IRS1), protein kinase-B (PKB) and extracellular regulated protein kinase (ERK1/ERK2). There was no significant difference i...
Source: BJ Disease - December 4, 2008 Category: Biochemistry Authors: M Rajkhowa, S Brett, D J Cuthbertson, C Lipina, A J Ruiz-Alcaraz, G E Thomas, L Logie, J R Petrie, C Sutherland Tags: BJ Signal Source Type: journals