Archaeal Mo-Containing Glyceraldehyde Oxidoreductase Isozymes Exhibit Diverse Substrate Specificities through Unique Subunit Assemblies

Archaea use glycolytic pathways distinct from those found in bacteria and eukaryotes, where unique enzymes catalyze each reaction step. In this study, we isolated three isozymes of glyceraldehyde oxidoreductase (GAOR1, GAOR2 and GAOR3) from the thermoacidophilic archaeon Sulfolobus tokodaii. GAOR1–3 belong to the xanthine oxidoreductase superfamily, and are composed of a molybdo-pyranopterin subunit (L), a flavin subunit (M), and an iron-sulfur subunit (S), forming an LMS hetero-trimer unit. We found that GAOR1 is a tetramer of the {"type":"entrez-protein","attrs":{"text":"STK17810","term_id":"1438759421"}}STK17810/{"type":"entrez-protein","attrs":{"text":"STK17830","term_id":"1438888158"}}STK17830/{"type":"entrez-protein","attrs":{"text":"STK17820","term_id":"1438888149"}}STK17820 hetero-trimer, GAOR2 is a dimer of the {"type":"entrez-protein","attrs":{"text":"STK23390","term_id":"1439642066"}}STK23390/{"type":"entrez-protein","attrs":{"text":"STK05620","term_id":"1438918159"}}STK05620/{"type":"entrez-protein","attrs":{"text":"STK05610","term_id":"1439044704"}}STK05610 hetero-trimer, and GAOR3 is the {"type":"entrez-protein","attrs":{"text":"STK24840","term_id":"1439962493"}}STK24840/{"type":"entrez-protein","attrs":{"text":"STK05620","term_id":"1438918159"}}STK05620/{"type":"entrez-protein","attrs":{"text":"STK05610","term_id":"1439044704"}}STK05610 hetero-trimer. GAOR1–3 exhibited diverse substrate specificities for their electron donors and acceptors, due to their different L-subunits, and probably participate in the non-phosphorylative Entner-Doudoroff glycolytic pathway. We determined the crystal structure of GAOR2, as the first three-dimensional structure of an archaeal molybdenum-containing hydroxylase, to obtain structural insights into their substrate specificities and subunit assemblies. The gene arrangement and the crystal structure suggested that the M/S-complex serves as a structural scaffold for the binding of the L-subunit, to construct the three enzymes with different specificities. Collectively, our findings illustrate a novel principle of a prokaryotic multicomponent isozyme system.     

Modulation of Platelet Activation and Thrombus Formation Using a Pan-PI3K Inhibitor S14161

The phosphatidylinositol 3–kinase (PI3K) signaling pathway is critical in modulating platelet functions. In the present study, we evaluated the effect of {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161, a recently identified pan-class I PI3K inhibitor, on platelet activation and thrombus formation. Results showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 inhibited human platelet aggregation induced by collagen, thrombin, U46619, and ADP in a dose-dependent manner. Flow cytometric studies showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 inhibited convulxin- or thrombin-induced P-selectin expression and fibrinogen binding of single platelet. {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 also inhibited platelet spreading on fibrinogen and clot retraction, processes mediated by outside-in signaling. Using a microfluidic chamber we demonstrated that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 decreased platelet adhesion on collagen-coated surface by about 80%. Western blot showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 inhibited phosphorylation of Akt at both Ser473 and Thr308 sites, and GSK3β at Ser9 in response to collagen, thrombin, or U46619. Comparable studies showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 has a higher potential bioavailability than {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002, a prototypical inhibitor of pan-class I PI3K. Finally, the effects of {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 on thrombus formation in vivo were measured using a ferric chloride-induced carotid artery injury model in mice. The intraperitoneal injection of {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 (2 mg/kg) to male C57BL/6 mice significantly extended the first occlusion time (5.05±0.99 min, n = 9) compared to the vehicle controls (3.72±0.95 min, n = 8) (P<0.05), but did not prolong the bleeding time (P>0.05). Taken together, our data showed that {"type":"entrez-protein","attrs":{"text":"S14161","term_id":"98844","term_text":"pir||S14161"}}S14161 inhibits platelet activation and thrombus formation without significant bleeding tendency and toxicity, and considering its potential higher bioavailability, it may be developed as a novel therapeutic agent for the prevention of thrombotic disorders.     

Allosteric Modulation of a Cannabinoid G Protein-coupled Receptor: BINDING SITE ELUCIDATION AND RELATIONSHIP TO G PROTEIN SIGNALING*

The cannabinoid 1 (CB₁) allosteric modulator, 5-chloro-3-ethyl-1H-indole-2-carboxylic acid [2-(4-piperidin-1-yl-phenyl)-ethyl]-amide) ({"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569), has the paradoxical effect of increasing the equilibrium binding of [³H](−)-3-[2-hydroxyl-4-(1,1-dimethylheptyl)phenyl]-4-[3-hydroxylpropyl]cyclohexan-1-ol (CP55,940, an orthosteric agonist) while at the same time decreasing its efficacy (in G protein-mediated signaling). {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 also decreases basal signaling, acting as an inverse agonist for the G protein-mediated signaling pathway. In ligand displacement assays, {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 can displace the CB₁ antagonist/inverse agonist, N-(piperidiny-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide(SR141716A). The goal of this work was to identify the binding site of {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 at CB₁. To this end, we used computation, synthesis, mutation, and functional studies to identify the {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569-binding site in the CB₁ TMH3-6-7 region. This site is consistent with the results of K3.28¹⁹²A, F3.36²⁰⁰A, W5.43²⁷⁹A, W6.48³⁵⁶A, and F3.25¹⁸⁹A mutation studies, which revealed the {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569-binding site overlaps with our previously determined binding site of SR141716A but extends extracellularly. Additionally, we identified a key electrostatic interaction between the {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 piperidine ring nitrogen and K3.28¹⁹² that is important for {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 to act as an inverse agonist. At this allosteric site, {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 promotes an intermediate conformation of the CB₁ receptor, explaining {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569''s ability to increase equilibrium binding of CP55,940. This site also explains {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569''s ability to antagonize the efficacy of CP55,940 in three complementary ways. 1) {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 sterically blocks movements of the second extracellular loop that have been linked to receptor activation. 2) {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 sterically blocks a key electrostatic interaction between the third extracellular loop residue Lys-373 and D2.63¹⁷⁶. 3) {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 packs against TMH6, sterically hindering movements of this helix that have been shown to be important for receptor activation.     

BRCA2 is needed for both repair and cell cycle arrest in mammalian cells exposed to S23906, an anticancer monofunctional DNA binder

Repair of DNA-targeted anticancer agents is an active area of investigation of both fundamental and clinical interest. However, most studies have focused on a small number of compounds limiting our understanding of both DNA repair and the DNA damage response. {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 is an acronycine derivative that shows strong activity toward solid tumors in experimental models. {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 forms bulky monofunctional DNA adducts in the minor groove which leads to destabilization of the double-stranded helix. We now report that {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 induces formation of DNA double strand breaks that are processed through homologous recombination (HR) but not Non-Homologous End-Joining (NHEJ) repair. Interestingly, {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 exposure was accompanied by a higher sensitivity of BRCA2-deficient cells compared to other HR deficient cell lines and by an S-phase accumulation in wild-type (wt), but not in BRCA2-deficient cells. Recently, we have shown that {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906-induced S phase arrest was mediated by the checkpoint kinase Chk1. However, its activated phosphorylated form is equally induced by {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 in wt and BRCA2-deficient cells, likely indicating a role for BRCA2 downstream of Chk1. Accordingly, override of the S phase arrest by either 7-hydroxystaurosporine (UCN-01) or AZD7762 potentiates the cytotoxic activity of {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 in wt, but not in BRCA2-deficient cells. Together, our findings suggest that the pronounced sensitivity of BRCA2-deficient cells to {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906 is due to both a defective S-phase arrest and the absence of HR repair. Tumors with deficiencies for proteins involved in HR, and BRCA2 in particular, may thus show increased sensitivity to {"type":"entrez-protein","attrs":{"text":"S23906","term_id":"96914","term_text":"pir||S23906"}}S23906, thereby providing a rationale for patient selection in clinical trials.     

Distinct Roles of β-Arrestin 1 and β-Arrestin 2 in ORG27569-induced Biased Signaling and Internalization of the Cannabinoid Receptor 1 (CB1)

The cannabinoid receptor 1 (CB1) is a G protein-coupled receptor primarily expressed in brain tissue that has been implicated in several disease states. CB1 allosteric compounds, such as {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569, offer enormous potential as drugs over orthosteric ligands, but their mechanistic, structural, and downstream effects upon receptor binding have not been established. Previously, we showed that {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 enhances agonist binding affinity to CB1 but inhibits G protein-dependent agonist signaling efficacy in HEK293 cells and rat brain expressing the CB1 receptor (Ahn, K. H., Mahmoud, M. M., and Kendall, D. A. (2012) J. Biol. Chem. 287, 12070–12082). Here, we identify the mediators of CB1 receptor internalization and {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569-induced G protein-independent signaling. Using siRNA technology, we elucidate an {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569-induced signaling mechanism for CB1 wherein β-arrestin 1 mediates short term signaling to ERK1/2 with a peak at 5 min and other upstream kinase components including MEK1/2 and c-Src. Consistent with these findings, we demonstrate co-localization of CB1-GFP with red fluorescent protein-β-arrestin 1 upon {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569 treatment using confocal microscopy. In contrast, we show the critical role of β-arrestin 2 in CB1 receptor internalization upon treatment with CP55940 (agonist) or treatment with {"type":"entrez-protein","attrs":{"text":"ORG27569","term_id":"1179174593","term_text":"ORG27569"}}ORG27569. These results demonstrate for the first time the involvement of β-arrestin in CB1-biased signaling by a CB1 allosteric modulator and also define the differential role of the two β-arrestin isoforms in CB1 signaling and internalization.     

Analysis of the Arabinose-5-Phosphate Isomerase of Bacteroides fragilis Provides Insight into Regulation of Single-Domain Arabinose Phosphate Isomerases

Arabinose-5-phosphate isomerases (APIs) catalyze the interconversion of d-ribulose-5-phosphate and d-arabinose-5-phosphate, the first step in the biosynthesis of 3-deoxy-d-manno-octulosonic acid (Kdo), an essential component of the lipopolysaccharide in Gram-negative bacteria. Classical APIs, such as Escherichia coli KdsD, contain a sugar isomerase domain and a tandem cystathionine beta-synthase domain. Despite substantial effort, little is known about structure-function relationships in these APIs. We recently reported an API containing only a sugar isomerase domain. This protein, c3406 from E. coli CFT073, has no known physiological function. In this study, we investigated a putative single-domain API from the anaerobic Gram-negative bacterium Bacteroides fragilis. This putative API (UniProt ID {"type":"entrez-protein","attrs":{"text":"Q5LIW1","term_id":"81317194","term_text":"Q5LIW1"}}Q5LIW1) is the only protein encoded by the B. fragilis genome with significant identity to any known API, suggesting that it is responsible for lipopolysaccharide biosynthesis in B. fragilis. We tested this hypothesis by preparing recombinant {"type":"entrez-protein","attrs":{"text":"Q5LIW1","term_id":"81317194","term_text":"Q5LIW1"}}Q5LIW1 protein (here referred to by the UniProt ID {"type":"entrez-protein","attrs":{"text":"Q5LIW1","term_id":"81317194","term_text":"Q5LIW1"}}Q5LIW1), characterizing its API activity in vitro, and demonstrating that the gene encoding {"type":"entrez-protein","attrs":{"text":"Q5LIW1","term_id":"81317194","term_text":"Q5LIW1"}}Q5LIW1 (GenBank ID {"type":"entrez-protein","attrs":{"text":"YP_209877.1","term_id":"60679733","term_text":"YP_209877.1"}}YP_209877.1) was able to complement an API-deficient E. coli strain. We demonstrated that {"type":"entrez-protein","attrs":{"text":"Q5LIW1","term_id":"81317194","term_text":"Q5LIW1"}}Q5LIW1 is inhibited by cytidine 5′-monophospho-3-deoxy-d-manno-2-octulosonic acid, the final product of the Kdo biosynthesis pathway, with a K_i of 1.91 μM. These results support the assertion that {"type":"entrez-protein","attrs":{"text":"Q5LIW1","term_id":"81317194","term_text":"Q5LIW1"}}Q5LIW1 is the API that supports lipopolysaccharide biosynthesis in B. fragilis and is subject to feedback regulation by CMP-Kdo. The sugar isomerase domain of E. coli KdsD, lacking the two cystathionine beta-synthase domains, demonstrated API activity and was further characterized. These results suggest that {"type":"entrez-protein","attrs":{"text":"Q5LIW1","term_id":"81317194","term_text":"Q5LIW1"}}Q5LIW1 may be a suitable system to study API structure-function relationships.     

Electrophysiological Effects of SKF83959 on Hippocampal CA1 Pyramidal Neurons: Potential Mechanisms for the Drug's Neuroprotective Effects

Although the potent anti-parkinsonian action of the atypical D₁-like receptor agonist {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 has been attributed to the selective activation of phosphoinositol(PI)-linked D₁ receptor, whereas the mechanism underlying its potent neuroprotective effect is not fully understood. In the present study, the actions of {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 on neuronal membrane potential and neuronal excitability were investigated in CA1 pyramidal neurons of rat hippocampal slices. {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 (10–100 µM) caused a concentration-dependent depolarization, associated with a reduction of input resistance in CA1 pyramidal neurons. The depolarization was blocked neither by antagonists for D₁, D₂, 5-HT_2A/2C receptors and α₁-adrenoceptor, nor by intracellular dialysis of GDP-β-S. However, the specific HCN channel blocker ZD7288 (10 µM) antagonized both the depolarization and reduction of input resistance caused by {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959. In voltage-clamp experiments, {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 (10–100 µM) caused a concentration-dependent increase of Ih current in CA1 pyramidal neurons, which was independent of D₁ receptor activation. Moreover, {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 (50 µM) caused a 6 mV positive shift in the activation curve of Ih and significantly accelerated the activation of Ih current. In addition, {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 also reduced the neuronal excitability of CA1 pyramidal neurons, which was manifested by the decrease in the number and amplitude of action potentials evoked by depolarizing currents, and by the increase of firing threshold and rhoebase current. The above results suggest that {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 increased Ih current through a D₁ receptor-independent mechanism, which led to the depolarization of hippocampal CA1 pyramidal neurons. These findings provide a novel mechanism for the drug''s neuroprotective effects, which may contributes to its therapeutic benefits in Parkinson''s disease.     

Aminostyrylbenzofuran Directly Reduces Oligomeric Amyloid-β and Reverses Cognitive Deficits in Alzheimer Transgenic Mice

Alzheimer''s disease is an irreversible neurodegenerative disorder that is characterized by the abnormal aggregation of amyloid-β into neurotoxic oligomers and plaques. Although many disease-modifying molecules are currently in Alzheimer clinical trials, a small molecule that inhibits amyloid-β aggregation and ameliorates the disorder has not been approved to date. Herein, we report the effects of a potent small molecule, 6-methoxy-2-(4-dimethylaminostyryl) benzofuran ({"type":"entrez-protein","attrs":{"text":"KMS88009","term_id":"870800947","term_text":"KMS88009"}}KMS88009), that directly disrupts amyloid-β oligomerization, preserving cognitive behavior when used prophylactically and reversing declines in cognitive behavior when used therapeutically. {"type":"entrez-protein","attrs":{"text":"KMS88009","term_id":"870800947","term_text":"KMS88009"}}KMS88009 exhibited excellent pharmacokinetic profiles with extensive brain uptake and a high level of safety. When orally administered before and after the onset of Alzheimer''s disease symptoms, {"type":"entrez-protein","attrs":{"text":"KMS88009","term_id":"870800947","term_text":"KMS88009"}}KMS88009 significantly reduced assembly of amyloid-β oligomers and improved cognitive behaviors in the APP/PS1 double transgenic mouse model. The unique dual mode of action indicates that {"type":"entrez-protein","attrs":{"text":"KMS88009","term_id":"870800947","term_text":"KMS88009"}}KMS88009 may be a powerful therapeutic candidate for the treatment of Alzheimer''s disease.     

Arylbenzazepines Are Potent Modulators for the Delayed Rectifier K+ Channel: A Potential Mechanism for Their Neuroprotective Effects

(±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959, like many other arylbenzazepines, elicits powerful neuroprotection in vitro and in vivo. The neuroprotective action of the compound was found to partially depend on its D₁-like dopamine receptor agonistic activity. The precise mechanism for the (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959-mediated neuroprotection remains elusive. We report here that (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 is a potent blocker for delayed rectifier K⁺ channel. (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 inhibited the delayed rectifier K⁺ current (I _K) dose-dependently in rat hippocampal neurons. The IC ₅₀ value for inhibition of I _K was 41.9±2.3 µM (Hill coefficient = 1.81±0.13, n = 6), whereas that for inhibition of I _A was 307.9±38.5 µM (Hill coefficient = 1.37±0.08, n = 6). Thus, (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 is 7.3-fold more potent in suppressing I _K than I _A. Moreover, the inhibition of I _K by (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 was voltage-dependent and not related to dopamine receptors. The rapidly onset of inhibition and recovery suggests that the inhibition resulted from a direct interaction of (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 with the K⁺ channel. The intracellular application of (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 had no effects of on I _K, indicating that the compound most likely acts at the outer mouth of the pore of K⁺ channel. We also tested the enantiomers of (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959, R-(+) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 (MCL-201), and S-(−) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959 (MCL-202), as well as {"type":"entrez-protein","attrs":{"text":"SKF38393","term_id":"1157151916","term_text":"SKF38393"}}SKF38393; all these compounds inhibited I _K. However, (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959, at either 0.1 or 1 mM, exhibited the strongest inhibition on the currents among all tested drug. The present findings not only revealed a new potent blocker of I _K , but also provided a novel mechanism for the neuroprotective action of arylbenzazepines such as (±) {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959.     

Efficacy of neuraminidase (NA) inhibitors against H1N1 strains of different geographical regions: an in silico approach

This report described the efficacy of NA inhibitors against newly evolved strains of H1N1 viruses. This in silico study was designed to understand the mode of interactions of NA inhibitors with NA. Hence, ligand, oseltamivir, zanamivir and peramivir were docked with modeled NA, {"type":"entrez-protein","attrs":{"text":"ACD65204","term_id":"188572592","term_text":"ACD65204"}}ACD65204 (USA/2007), {"type":"entrez-protein","attrs":{"text":"BAA06717","term_id":"1212736","term_text":"BAA06717"}}BAA06717 (Japan/1992), {"type":"entrez-protein","attrs":{"text":"ACE77988","term_id":"190403802","term_text":"ACE77988"}}ACE77988 (S. Korea/2005) and {"type":"entrez-protein","attrs":{"text":"ACD65204","term_id":"188572592","term_text":"ACD65204"}}ACD65204 (USA/2007). This study is based on interaction energies. Ramachandran Z-scores for these modeled structures were found to be −0.998, −1.121, −0.870 and −1.023, respectively, which confirms the accuracy of the modeled structures. These interactions revealed that some of these interacting residues have remained conserved throughout all the pandemics. These amino acid residues were found to be R118, R152, R225, E277, E278, R293 and Y402. Moreover, our study concludes that peramivir is the most efficient inhibitor against NA of H1N1.     

Role for the A Domain of Unprocessed Accumulation-Associated Protein (Aap) in the Attachment Phase of the Staphylococcus epidermidis Biofilm Phenotype

The polysaccharide intercellular adhesin or the cell wall-anchored accumulation-associated protein (Aap) mediates cellular accumulation during Staphylococcus epidermidis biofilm maturation. Mutation of sortase, which anchors up to 11 proteins (including Aap) to the cell wall, blocked biofilm development by the cerebrospinal fluid isolate {"type":"entrez-protein","attrs":{"text":"CSF41498","term_id":"904025233","term_text":"CSF41498"}}CSF41498. Aap was implicated in this phenotype when Western blots and two-dimensional (2D) electrophoresis revealed increased levels of the protein in culture supernatants. Unexpectedly, reduced levels of primary attachment were associated with impaired biofilm formation by {"type":"entrez-protein","attrs":{"text":"CSF41498","term_id":"904025233","term_text":"CSF41498"}}CSF41498 srtA and aap mutants. In contrast to previous studies, which implicated Aap proteolytic cleavage and, specifically, the Aap B domains in biofilm accumulation, the {"type":"entrez-protein","attrs":{"text":"CSF41498","term_id":"904025233","term_text":"CSF41498"}}CSF41498 Aap protein was unprocessed. Furthermore, aap appeared to play a less important role in the biofilm phenotype of S. epidermidis 1457, in which the Aap protein is processed. Anti-Aap A-domain IgG inhibited primary attachment and biofilm formation in strain {"type":"entrez-protein","attrs":{"text":"CSF41498","term_id":"904025233","term_text":"CSF41498"}}CSF41498 but not in strain 1457. The nucleotide sequences of the aap gene A-domain region and cleavage site in strains {"type":"entrez-protein","attrs":{"text":"CSF41498","term_id":"904025233","term_text":"CSF41498"}}CSF41498 and 1457 were identical, implicating altered protease activity in the differential Aap processing results in the two strains. These data reveal a new role for the A domain of unprocessed Aap in the attachment phase of biofilm formation and suggest that extracellular protease activity can influence whether Aap contributes to the attachment or accumulation phases of the S. epidermidis biofilm phenotype.     

Modeling and structural analysis of cellulases using Clostridium thermocellum as template

Cellulase is one of the most widely distributed enzymes with wide application. They are involved in conversion of biomass into simpler sugars. Cellulase of Trichoderma longibrachiatum, a known cellulolytic fungus was compared with Clostridium thermocellum [{"type":"entrez-protein","attrs":{"text":"AAA23226.1","term_id":"289859","term_text":"AAA23226.1"}}AAA23226.1] cellulase. Blastp was performed with {"type":"entrez-protein","attrs":{"text":"AAA23226.1","term_id":"289859","term_text":"AAA23226.1"}}AAA23226.1 as query sequence to obtain nine similar sequences from NCBI protein data bank. The physicochemical properties of cellulase were analyzed using ExPASy’s ProtParam tool namely ProtParam, SOPMA and GOR IV. Homology modeling was done using SWISS MODEL and checked quality by RMSD values using VMD1.9.1. Active sites of each model were predicted using automated active site prediction server of SCFBio. Study revealed instability of cellulase of two eukaryotic strains namely Trichoderma longibrachiatum [{"type":"entrez-protein","attrs":{"text":"CAA43059.1","term_id":"5182","term_text":"CAA43059.1"}}CAA43059.1] and Melanocarpus albomyces [{"type":"entrez-protein","attrs":{"text":"CAD56665.1","term_id":"27125829","term_text":"CAD56665.1"}}CAD56665.1]. The negative GRAVY score value of cellulases ensured better interaction and activity in aqueous phase. It was found that molecular weight (M. Wt) ranges between 25-127.56 kDa. Iso-electric point (pI) of cellulases was found to be acidic in nature. GOR IV and SOPMA were used to predict secondary structure of cellulase, which showed that random coil, was dominated. Neighbor joining tree with C. thermocellum [{"type":"entrez-protein","attrs":{"text":"AAA23226.1","term_id":"289859","term_text":"AAA23226.1"}}AAA23226.1] cellulase as root showed that cellulases of Thermoaerobacter subterraneus [{"type":"entrez-protein","attrs":{"text":"ZP_07835928","term_id":"313902527","term_text":"ZP_07835928"}}ZP_07835928] and C. thermocellum [CAA4305.1] were more similar to eukaryotic cellulases supported by least boot strap values. Pseudoalteromonas haloplanktis cellulase was found to be the ideal model supported by least RMSD score among the predicted structures. Trichoderma longibrachiatum cellulase was found to be the best compared to other cellulases, which possess high number of active sites with ASN and THR rich active sites. CYS residues were also present ensuring stable interaction and better bonding. Hydrophilic residues were found high in active sites of all analyzed models and template.     

CGP37157, an inhibitor of the mitochondrial Na+/Ca2+ exchanger,protects neurons from excitotoxicity by blocking voltage-gated Ca2+ channels

Inhibition of the mitochondrial Na⁺/Ca²⁺ exchanger (NCLX) by {"type":"entrez-protein","attrs":{"text":"CGP37157","term_id":"875406365","term_text":"CGP37157"}}CGP37157 is protective in models of neuronal injury that involve disruption of intracellular Ca²⁺ homeostasis. However, the Ca²⁺ signaling pathways and stores underlying neuroprotection by that inhibitor are not well defined. In the present study, we analyzed how intracellular Ca²⁺ levels are modulated by {"type":"entrez-protein","attrs":{"text":"CGP37157","term_id":"875406365","term_text":"CGP37157"}}CGP37157 (10 μM) during NMDA insults in primary cultures of rat cortical neurons. We initially assessed the presence of NCLX in mitochondria of cultured neurons by immunolabeling, and subsequently, we analyzed the effects of {"type":"entrez-protein","attrs":{"text":"CGP37157","term_id":"875406365","term_text":"CGP37157"}}CGP37157 on neuronal Ca²⁺ homeostasis using cameleon-based mitochondrial Ca²⁺ and cytosolic Ca²⁺ ([Ca²⁺]_i) live imaging. We observed that NCLX-driven mitochondrial Ca²⁺ exchange occurs in cortical neurons under basal conditions as {"type":"entrez-protein","attrs":{"text":"CGP37157","term_id":"875406365","term_text":"CGP37157"}}CGP37157 induced a decrease in [Ca²]_i concomitant with a Ca²⁺ accumulation inside the mitochondria. In turn, {"type":"entrez-protein","attrs":{"text":"CGP37157","term_id":"875406365","term_text":"CGP37157"}}CGP37157 also inhibited mitochondrial Ca²⁺ efflux after the stimulation of acetylcholine receptors. In contrast, {"type":"entrez-protein","attrs":{"text":"CGP37157","term_id":"875406365","term_text":"CGP37157"}}CGP37157 strongly prevented depolarization-induced [Ca²⁺]_i increase by blocking voltage-gated Ca²⁺ channels (VGCCs), whereas it did not induce depletion of ER Ca²⁺ stores. Moreover, mitochondrial Ca²⁺ overload was reduced as a consequence of diminished Ca²⁺ entry through VGCCs. The decrease in cytosolic and mitochondrial Ca²⁺ overload by {"type":"entrez-protein","attrs":{"text":"CGP37157","term_id":"875406365","term_text":"CGP37157"}}CGP37157 resulted in a reduction of excitotoxic mitochondrial damage, characterized here by a reduction in mitochondrial membrane depolarization, oxidative stress and calpain activation. In summary, our results provide evidence that during excitotoxicity {"type":"entrez-protein","attrs":{"text":"CGP37157","term_id":"875406365","term_text":"CGP37157"}}CGP37157 modulates cytosolic and mitochondrial Ca²⁺ dynamics that leads to attenuation of NMDA-induced mitochondrial dysfunction and neuronal cell death by blocking VGCCs.     

Genome sequences published outside of Standards in Genomic Sciences, October �C November 2011

The purpose of this table is to provide the community with a citable record of publications of ongoing genome sequencing projects that have led to a publication in the scientific literature. While our goal is to make the list complete, there is no guarantee that we may have omitted one or more publications appearing in this time frame. Readers and authors who wish to have publications added to subsequent versions of this list are invited to provide the bibliographic data for such references to the SIGS editorial office.

• Phylum Crenarchaeota
• Thermoproteus tenax, strain Kra1, DSM 2078^T sequence accession {"type":"entrez-nucleotide","attrs":{"text":"FN869859","term_id":"350274033"}}FN869859 [1]
• Phylum Euryarchaeota
• Haloarcula hispanica CGMCC 1.2049, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002921","term_id":"343781660"}}CP002921 (chromosome I), {"type":"entrez-nucleotide","attrs":{"text":"CP002922","term_id":"343784702"}}CP002922 (chromosome II), and {"type":"entrez-nucleotide","attrs":{"text":"CP002923","term_id":"343785159"}}CP002923 (plasmid pHH400) [2]
• Methanococcus maripaludis, strain X1 (unculturable) sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002913","term_id":"339903450"}}CP002913 [3]
• Phylum Proteobacteria
• Acinetobacter baumannii strain 1656-2, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP001921","term_id":"322506180"}}CP001921 [4]
• Arcobacter butzleri strain ED-1, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AP012047","term_id":"345468266"}}AP012047, {"type":"entrez-nucleotide","attrs":{"text":"AP012048","term_id":"345470425"}}AP012048, and {"type":"entrez-nucleotide","attrs":{"text":"AP012049","term_id":"345473271"}}AP012049 [5]
• Brucella suis strain 1330, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002997","term_id":"343381878"}}CP002997 and {"type":"entrez-nucleotide","attrs":{"text":"CP002998","term_id":"343384001"}}CP002998 [6]
• Campylobacter fetus subsp. venerealis NCTC 10354, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFGH01000000","term_id":"342326654"}}AFGH01000000 [7]
• “Chromobacterium sp.” strain C-61, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CAEE01000001","term_id":"344197715"}}CAEE01000001 to {"type":"entrez-nucleotide","attrs":{"text":"CAEE01001118","term_id":"344196598"}}CAEE01001118 [8]
• Cronobacter sakazakii strain E899, sequence accession {"type":"entrez-protein","attrs":{"text":"AFMO00000000","term_id":"333956885"}}AFMO00000000 [9]
• “Desulfovibrio sp.” strain A2, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGFG01000000","term_id":"347520603"}}AGFG01000000 [10]
• “Erythrobacter sp.” strain NAP1, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"NZ_AAMW00000000","term_id":"85710750"}}NZ_AAMW00000000 [11]
• Escherichia coli strain XH140A, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFVX01000000","term_id":"342364931"}}AFVX01000000 [12]
• Escherichia coli strain XH001, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFYG01000000","term_id":"344195679"}}AFYG01000000 [13]
• Haemophilus haemolyticus strain {"type":"entrez-nucleotide","attrs":{"text":"M19107","term_id":"1059793989"}}M19107, sequence accession {"type":"entrez-protein","attrs":{"text":"AFQN00000000","term_id":"341956667"}}AFQN00000000 [14]
• Haemophilus haemolyticus strain {"type":"entrez-nucleotide","attrs":{"text":"M19501","term_id":"147422"}}M19501, sequence accession {"type":"entrez-protein","attrs":{"text":"AFQO00000000","term_id":"341951174"}}AFQO00000000 [14]
• Haemophilus haemolyticus strain {"type":"entrez-nucleotide","attrs":{"text":"M21127","term_id":"1015602694"}}M21127, sequence accession {"type":"entrez-protein","attrs":{"text":"AFQP00000000","term_id":"341951709"}}AFQP00000000 [14]
• Haemophilus haemolyticus strain {"type":"entrez-nucleotide","attrs":{"text":"M21621","term_id":"189206"}}M21621, sequence accession {"type":"entrez-protein","attrs":{"text":"AFQQ00000000","term_id":"341955662"}}AFQQ00000000 [14]
• Haemophilus haemolyticus strain {"type":"entrez-nucleotide","attrs":{"text":"M21639","term_id":"1036032350"}}M21639, sequence accession {"type":"entrez-protein","attrs":{"text":"AFQR00000000","term_id":"341956942"}}AFQR00000000 [14]
• Idiomarina sp.” strain A28L, sequence accession {"type":"entrez-nucleotide-range","attrs":{"text":"AFPO01000001 to AFPO01000028","start_term":"AFPO01000001","end_term":"AFPO01000028","start_term_id":"336283061","end_term_id":"336280888"}}AFPO01000001 to AFPO01000028 [15]
• Ketogulonicigenium vulgare” strain WSH-001, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002018","term_id":"343461404"}}CP002018 (chromosome), {"type":"entrez-nucleotide","attrs":{"text":"CP002019","term_id":"343464001"}}CP002019 (plasmid pKVU_100), and {"type":"entrez-nucleotide","attrs":{"text":"CP002020","term_id":"343464245"}}CP002020 (plasmid pKVU_200) [16]
• Methylobacter tundripaludum strain SV96, sequence accession {"type":"entrez-protein","attrs":{"text":"AEGW00000000","term_id":"344263587"}}AEGW00000000 [17]
• Pseudogulbenkiania sp.” strain NH8B, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AP012224","term_id":"345641016"}}AP012224 [18]
• Pseudomonas aeruginosa NCGM1179, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"DF126593","term_id":"346055306"}}DF126593 through {"type":"entrez-nucleotide","attrs":{"text":"DF126613","term_id":"346061544"}}DF126613 [19]
• Pseudomonas putida strain B001, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CAED01000001","term_id":"344196578"}}CAED01000001 to {"type":"entrez-nucleotide","attrs":{"text":"CAEE01000262","term_id":"344197454"}}CAEE01000262 [20]
• Pseudomonas putida strain B6-2, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGCS01000000","term_id":"1026254431"}}AGCS01000000 [21]
• Pseudomonas stutzeri CGMCC 1.1803, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002881","term_id":"338799449"}}CP002881 [22]
• Ralstonia solanacearum phylotype IB, strain Y45, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFWL01000000","term_id":"343176764"}}AFWL01000000 [23]
• Rheinheimera sp.” strain A13L, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFHI01000001","term_id":"335881665"}}AFHI01000001 through {"type":"entrez-nucleotide","attrs":{"text":"AFHI01000072","term_id":"335877953"}}AFHI01000072 [24]
• Sphingobium yanoikuyae strain XLDN2-5, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFXE01000000","term_id":"345099327"}}AFXE01000000 [25]
• Vibrio cholerae strain Amazonia, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFSV01000000","term_id":"341584524"}}AFSV01000000 [26]
• Phylum Firmicutes
• Bacillus coagulans strain XZL4, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFWM01000000","term_id":"343381451"}}AFWM01000000 [27]
• Bacillus megaterium strain WSH-002, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP003017","term_id":"345441867"}}CP003017 (chromosome), plasmids {"type":"entrez-nucleotide","attrs":{"text":"CP003018","term_id":"345447048"}}CP003018 (plasmid pBME_100), {"type":"entrez-nucleotide","attrs":{"text":"CP003019","term_id":"345447118"}}CP003019 (plasmid pBME_200), and {"type":"entrez-nucleotide","attrs":{"text":"CP003020","term_id":"345447130"}}CP003020 (plasmid pBME_300) [28]
• Bacillus pumilus strain S-1, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGBY00000000","term_id":"346722060"}}AGBY00000000 [29]
• “Desulfosporosinus sp.” strain OT, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGAF01000000","term_id":"344330753"}}AGAF01000000 [30]
• Lentibacillus jeotgali strain Grbi, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGAV01000000","term_id":"345115115"}}AGAV01000000 [31]
• Leuconostoc carnosum KCTC 3525, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"BACM01000000","term_id":"338736686"}}BACM01000000 [32]
• Listeria ivanovii subsp. ivanovii strain PAM 55, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"FR687253","term_id":"346980581"}}FR687253 [33]
• Paenibacillus riograndensis strain SBR5, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGBD01000000","term_id":"345128217"}}AGBD01000000 [34]
• Sporolactobacillus inulinus strain CASD, sequence accession {"type":"entrez-protein","attrs":{"text":"AFVQ00000000","term_id":"823094357"}}AFVQ00000000 [35]
• Streptococcus pseudopneumoniae strain IS7493, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002925","term_id":"341932553"}}CP002925 and {"type":"entrez-nucleotide","attrs":{"text":"CP002926","term_id":"341934784"}}CP002926 [36]
• Streptococcus salivarius strain 57.I, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002888","term_id":"339291081"}}CP002888 and {"type":"entrez-nucleotide","attrs":{"text":"CP002889","term_id":"339293023"}}CP002889 [37]
• Streptococcus salivarius strain M18, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGBV01000000","term_id":"345528121"}}AGBV01000000 [38]
• Streptococcus suis SS12, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002640","term_id":"353733193"}}CP002640 [39]
• Streptococcus suis D9, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002641","term_id":"353735273"}}CP002641 [39]
• Streptococcus suis D12, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002644","term_id":"353737348"}}CP002644 [39]
• Streptococcus suis ST1, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002651","term_id":"353739427"}}CP002651 [39]
• Weissella thailandensis strain fsh4-2, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"HE575133","term_id":"341819811"}}HE575133 through {"type":"entrez-nucleotide","attrs":{"text":"HE575182","term_id":"341821297"}}HE575182 [40]
• Phylum Tenericutes
• Mycoplasma anatis strain 1340, sequence accession {"type":"entrez-protein","attrs":{"text":"AFVJ00000000","term_id":"341579585"}}AFVJ00000000 [41]
• Mycoplasma capricolum subsp. capripneumoniae strain M1601, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AENG01000000","term_id":"1067112011"}}AENG01000000 [42]
• Mycoplasma putrefaciens Type strain KS1, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP003021","term_id":"343956693"}}CP003021 [43]
• Corynebacterium pseudotuberculosis strain PAT10, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP002924","term_id":"341823823"}}CP002924 [44]
• Phylum Actinobacteria
• Bifidobacterium animalis subsp. lactis strain BLC1, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP003039","term_id":"447219159"}}CP003039 [45]
• Bifidobacterium breve strain DPC 6330, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFXX01000000","term_id":"347344070"}}AFXX01000000 [46]
• Brachybacterium squillarum strain M-6-3, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGBX01000000","term_id":"346317973"}}AGBX01000000 [47]
• “Citricoccus sp.” strain CH26A, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFXQ01000000","term_id":"344198218"}}AFXQ01000000 [48]
• Corynebacterium glutamicum strain S9114, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFYA01000000","term_id":"344046353"}}AFYA01000000 [49]
• Dietzia alimentaria strain 72, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGFF01000000","term_id":"347366987"}}AGFF01000000 [50]
• Mycobacterium colombiense CECT 3035, sequence accession {"type":"entrez-protein","attrs":{"text":"AFVW00000000","term_id":"400333858"}}AFVW00000000 [51]
• Mycobacterium tuberculosis NCGM2209, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"DF126614","term_id":"358229918"}}DF126614 and {"type":"entrez-nucleotide","attrs":{"text":"DF126615","term_id":"358233644"}}DF126615 [52]
• Rhodococcus erythropolis strain XP, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGCF01000000","term_id":"345896954"}}AGCF01000000 [53]
• Serinicoccus profundi MCCC 1A05965^T, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFYF00000000","term_id":"345107103"}}AFYF00000000 [54]
• Phylum Spirochaetes
• Leptospira interrogans, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP001221","term_id":"353456277"}}CP001221 (CI), {"type":"entrez-nucleotide","attrs":{"text":"CP001222","term_id":"353459699"}}CP001222 (CII) [55]
• Phylum Bacteroidetes
• Bacteroides faecis Type strain MAJ27^T, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AGDG01000000","term_id":"347014739"}}AGDG01000000 [56]
• Bizionia argentinensis, Type strain JUB59^T sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFXZ01000000","term_id":"344049519"}}AFXZ01000000 [57]
• Flavobacterium branchiophilum strain FL-15, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"FQ859183","term_id":"345528129"}}FQ859183 [58]
• “Flavobacteriaceae” strain S85, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"AFPK00000000","term_id":"339778866"}}AFPK00000000 [59]
• Phylum Thermotogae
• “Thermotoga sp.” strain RQ2, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"CP000969","term_id":"170175310"}}CP000969 [60]

Non-Bacterial genomes

• Aspergillus kawachii IFO 4308, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"DF126447","term_id":"358365263"}}DF126447 through {"type":"entrez-nucleotide","attrs":{"text":"DF126592","term_id":"358376882"}}DF126592, BACL01000001 through BACL01001641, {"type":"entrez-nucleotide","attrs":{"text":"AP012272","term_id":"354548779"}}AP012272 [61]
• Cajanus cajan pigeonpea, sequence accession PRJNA72815 [62]
• Coxsackievirus A22, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"JN542510","term_id":"354682107"}}JN542510 [63]
• Gordonia phage GRU1, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"JF923797","term_id":"356471393"}}JF923797 [64]
• Gordonia phage GTE5, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"JF923796","term_id":"356471300"}}JF923796 [64]
• Heterocephalus glaber naked mole rat, sequence accession {"type":"entrez-protein","attrs":{"text":"AFSB00000000","term_id":"351716052"}}AFSB00000000, {"type":"entrez-nucleotide","attrs":{"text":"AFSB01000000","term_id":"351581530"}}AFSB01000000 [65]
• Human Adenovirus Prototype 17, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"HQ910407","term_id":"324105303"}}HQ910407 [66]
• Macaca mulatta lasiota rhesus macaque, sequence accession {"type":"entrez-protein","attrs":{"text":"AEHL00000000","term_id":"355786683"}}AEHL00000000 [67]
• Macaca mulatta mulatta rhesus macaque, sequence accession {"type":"entrez-protein","attrs":{"text":"AEHK00000000","term_id":"355710505"}}AEHK00000000 [67]
• Porcine epidemic diarrhea virus, sequence accession {"type":"entrez-nucleotide","attrs":{"text":"JN547228","term_id":"351630105"}}JN547228 [68]

     

SKF83959, an Agonist of Phosphatidylinositol-Linked D1-Like Receptors,Promotes ERK1/2 Activation and Cell Migration in Cultured Rat Astrocytes

Extracellular signal-regulated kinase 1/2 (ERK1/2) is a member of the mitogen-activated protein kinase family. It can mediate cell migration. Classical dopamine receptor-mediated ERK1/2 phosphorylation is widely studied in neurons. Here, we report that ERK1/2 phosphorylation is also modulated by putative phosphatidylinositol-linked D₁-like receptors in cultured rat astrocytes. 6-chloro-7,8-dihydroxy-3-methyl-1-(3-methylphenyl)-2,3,4,5-tetrahydro-1H-3-benzazepine ({"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959), an agonist of the putative phosphatidylinositol-linked D₁-like receptors, was found to enhance ERK1/2 phosphorylation, which then promoted the migration of cultured astrocytes. The {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959-induced ERK1/2 phosphorylation was found to be Ca²⁺-independent based on the following observations: i. chelating intracellular Ca²⁺ did not inhibit ERK1/2 phosphorylation and astrocyte migration; ii. blockage of the release of intracellular Ca²⁺ from the endoplasmic reticulum by an inhibitor of inositol 1,4,5-trisphosphate (IP3) receptor did not attenuate ERK1/2 phosphorylation. However, inhibition of phospholipase C (PLC), the upstream molecule of internal Ca²⁺ release, disabled {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959’s ability to elevate the level of ERK1/2 phosphorylation. Both non-selective protein kinase C (PKC) inhibitor and PKCδ selective inhibitor prevented ERK1/2 phosphorylation increase and astrocyte migration, but PKCα inhibitor did not. This suggests that Ca²⁺-independent and diacylglycerol-dependent PKCδ acts downstream of putative phosphatidylinositol-linked D₁-like receptor activation and mediates {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959-induced elevation of ERK1/2 phosphorylation in order to modulate astrocyte migration. In conclusion, our results demonstrate that {"type":"entrez-protein","attrs":{"text":"SKF83959","term_id":"1155968032","term_text":"SKF83959"}}SKF83959-induced increases in ERK1/2 phosphorylation and astrocyte migration are dependent on PLC-PKCδ signals. This might help us to further understand the functions of the putative phosphatidylinositol-linked D₁-like receptors in the nervous system.     

Angiotensin II type 2 receptor correlates with therapeutic effects of losartan in rats with adjuvant‐induced arthritis

The angiotensin II type 1 receptor (AT1R) blocker losartan ameliorates rheumatoid arthritis (RA) in an experimental model. In RA, AT2R mainly opposes AT1R, but the mechanism by which this occurs still remains obscure. In the present study, we investigated the role of AT2R in the treatment of rats with adjuvant-induced arthritis (AIA) by losartan. Adjuvant-induced arthritis rats were treated with losartan (5, 10 and 15 mg/kg) and methotrexate (MTX; 0.5 mg/kg) in vivo from day 14 to day 28. Arthritis was evaluated by the arthritis index and histological examination. Angiotensin II, tumour necrosis factor-α, and VEGF levels were examined by ELISA. The expression of AT1R and AT2R was detected by western blot and immunohistochemistry analysis. After stimulation with interleukin-1β in vitro, the effects of the AT2R agonist {"type":"entrez-protein","attrs":{"text":"CGP42112","term_id":"874777115","term_text":"CGP42112"}}CGP42112 (10⁻⁸–10⁻⁵ M) on the chemotaxis of monocytes induced by 10% foetal calf serum (FCS) were analysed by using Transwell assay. Subsequently, the therapeutic effects of {"type":"entrez-protein","attrs":{"text":"CGP42112","term_id":"874777115","term_text":"CGP42112"}}CGP42112 (5, 10 and 20 μg/kg) were evaluated in vivo by intra-articular injection in AIA rats. After treatment with losartan, the down-regulation of AT1R expression and up-regulation of AT2R expression in the spleen and synovium of AIA rats correlated positively with reduction in the polyarthritis index. Treatment with {"type":"entrez-protein","attrs":{"text":"CGP42112","term_id":"874777115","term_text":"CGP42112"}}CGP42112 inhibited the chemotaxis of AIA monocytes in vitro, possibly because of the up-regulation of AT2R expression. Intra-articular injection with {"type":"entrez-protein","attrs":{"text":"CGP42112","term_id":"874777115","term_text":"CGP42112"}}CGP42112 (10 and 20 μg/kg) ameliorated the arthritis index and histological signs of arthritis. In summary, the present study strongly suggests that the up-regulation of AT2R might be an additional mechanism by which losartan exerts its therapeutic effects in AIA rats.