Second Messenger Metabolising Enzymes
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Recombinant Human Phosphodiesterase 4A/PDE4A
Recombinant Human Phosphodiesterase 4A/PDE4A Catalog Number: 7767-PE DESCRIPTION Source Spodoptera frugiperda, Sf 21 (baculovirus)derived Pro331Met723, with an Nterminal Met and a Cterminal 6His tag Accession # P27815 Nterminal Sequence Pro331 Analysis Predicted Molecular 46 kDa Mass SPECIFICATIONS SDSPAGE 4448 kDa, reducing conditions Activity Measured by its ability to convert cAMP to 5'AMP. The specific activity is >28,000 pmol/min/μg, as measured under the described conditions. Endotoxin Level <0.01 EU per 1 μg of the protein by the LAL method. Purity >95%, by SDSPAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane. Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl. See Certificate of Analysis for details. Activity Assay Protocol Materials l Assay Buffer (1X): 20 mM Tris, 1 mM MgCl2, 1 mM DTT, 0.01538% CHAPS, pH 7.5 l Recombinant Human Phosphodiesterase 4A/PDE4A (rhPDE4A) (Catalog # 7767PE) l Adenosine 3’,5’cyclic monophosphate (cAMP) (Sigma, Catalog # A6885) 0.1 M stock in deionized water l Sialyltransferase Activity Kit (Catalog # EA002) l 96well Clear Plate (Costar, Catalog # 92592) l Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent Assay 1. Dilute 1 mM Phosphate Standard provided by the Sialyltransferase Kit by adding 40 µL of the 1 mM Phosphate Standard to 360 µL of Assay Buffer for a 100 µM stock. 2. Continue standard curve by performing six additional onehalf serial dilutions of the 100 µM Phosphate stock in Assay Buffer. -
PDE4) Subtypes in Human Primary CD4+ T Cells: Predominant Role of PDE4D This Information Is Current As of September 26, 2021
Differential Expression and Function of Phosphodiesterase 4 (PDE4) Subtypes in Human Primary CD4+ T Cells: Predominant Role of PDE4D This information is current as of September 26, 2021. Daniel Peter, S. L. Catherine Jin, Marco Conti, Armin Hatzelmann and Christof Zitt J Immunol 2007; 178:4820-4831; ; doi: 10.4049/jimmunol.178.8.4820 http://www.jimmunol.org/content/178/8/4820 Downloaded from References This article cites 53 articles, 24 of which you can access for free at: http://www.jimmunol.org/content/178/8/4820.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 26, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2007 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Differential Expression and Function of Phosphodiesterase 4 :PDE4) Subtypes in Human Primary CD4؉ T Cells) Predominant Role of PDE4D1 Daniel Peter,* S. L. Catherine Jin,† Marco Conti,† Armin Hatzelmann,* and Christof Zitt2* Type 4 phosphodiesterases (PDE4) are critical regulators in TCR signaling by attenuating the negative constraint of cAMP. -
Reduced PDE4 Expression and Activity Contributes to Enhanced Catecholamine-Induced Camp Accumulation in Adipocytes from FOXC2 Transgenic Mice
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector FEBS Letters 580 (2006) 4126–4130 Reduced PDE4 expression and activity contributes to enhanced catecholamine-induced cAMP accumulation in adipocytes from FOXC2 transgenic mice Line M. Grønninga,*, George S. Baillieb, Anna Cederbergc, Martin J. Lynchb, Miles D. Houslayb, Sven Enerba¨ckc, Kjetil Taske´na a Biotechnology Centre of Oslo, University of Oslo, P.O. Box 1125 Blindern, 0317 Oslo, Norway b Dvn Biochemistry and Molecular Biology, IBLS, Wolfson Link Building, University of Glasgow, Glasgow G12 8QQ, Scotland, UK c Medical Genetics, Department of Medical Biochemistry, Go¨teborg University, SE 405 30 Go¨teborg, Sweden Received 11 April 2006; revised 14 June 2006; accepted 15 June 2006 Available online 30 June 2006 Edited by Laszlo Nagy diesterase (PDE) inhibitor IBMX to minimize hydrolysis of Abstract Overexpression of forkhead transcription factor FOXC2 in white adipose tissue (WAT) leads to a lean phenotype cAMP by PDEs. Thus, the strongly enhanced and sustained resistant to diet-induced obesity. This is due, in part, to enhanced cAMP response previously observed in FOXC2 transgenic catecholamine-induced cAMP-PKA signaling in FOXC2 trans- adipocytes is most likely a result of the increased expression genic mice. Here we show that rolipram treatment of adipocytes of b-AR receptors, since this would lead to a more profound from FOXC2 transgenic mice did not increase isoproterenol-in- activation of b-AR associated adenylyl cyclases (ACs) and, duced cAMP accumulation to the same extent as in wild type thus, increased generation of cAMP from ATP. -
Structure-Based Redesigning of Pentoxifylline Analogs Against
www.nature.com/scientificreports OPEN Structure‑based redesigning of pentoxifylline analogs against selective phosphodiesterases to modulate sperm functional competence for assisted reproductive technologies Mutyala Satish1,5, Sandhya Kumari2,5, Waghela Deeksha1, Suman Abhishek1, Kulhar Nitin1, Satish Kumar Adiga2, Padmaraj Hegde3, Jagadeesh Prasad Dasappa4, Guruprasad Kalthur2* & Eerappa Rajakumara1* Phosphodiesterase (PDE) inhibitors, such as pentoxifylline (PTX), are used as pharmacological agents to enhance sperm motility in assisted reproductive technology (ART), mainly to aid the selection of viable sperm in asthenozoospermic ejaculates and testicular spermatozoa, prior to intracytoplasmic sperm injection (ICSI). However, PTX is reported to induce premature acrosome reaction (AR) and, exert toxic efects on oocyte function and early embryo development. Additionally, in vitro binding studies as well as computational binding free energy (ΔGbind) suggest that PTX exhibits weak binding to sperm PDEs, indicating room for improvement. Aiming to reduce the adverse efects and to enhance the sperm motility, we designed and studied PTX analogues. Using structure‑guided in silico approach and by considering the physico‑chemical properties of the binding pocket of the PDEs, designed analogues of PTX. In silico assessments indicated that PTX analogues bind more tightly to PDEs and form stable complexes. Particularly, ex vivo evaluation of sperm treated with one of the PTX analogues (PTXm‑1), showed comparable benefcial efect at much lower concentration—slower -
Phosphodiesterase 1B Knock-Out Mice Exhibit Exaggerated Locomotor Hyperactivity and DARPP-32 Phosphorylation in Response to Dopa
The Journal of Neuroscience, June 15, 2002, 22(12):5188–5197 Phosphodiesterase 1B Knock-Out Mice Exhibit Exaggerated Locomotor Hyperactivity and DARPP-32 Phosphorylation in Response to Dopamine Agonists and Display Impaired Spatial Learning Tracy M. Reed,1,3 David R. Repaske,2* Gretchen L. Snyder,4 Paul Greengard,4 and Charles V. Vorhees1* Divisions of 1Developmental Biology and 2Endocrinology, Children’s Hospital Research Foundation, Cincinnati, Ohio 45229, 3Department of Biology, College of Mount St. Joseph, Cincinnati, Ohio 45233, and 4Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021 Using homologous recombination, we generated mice lack- maze spatial-learning deficits. These results indicate that en- ing phosphodiesterase-mediated (PDE1B) cyclic nucleotide- hancement of cyclic nucleotide signaling by inactivation of hydrolyzing activity. PDE1B Ϫ/Ϫ mice showed exaggerated PDE1B-mediated cyclic nucleotide hydrolysis plays a signifi- hyperactivity after acute D-methamphetamine administra- cant role in dopaminergic function through the DARPP-32 and tion. Striatal slices from PDE1B Ϫ/Ϫ mice exhibited increased related transduction pathways. levels of phospho-Thr 34 DARPP-32 and phospho-Ser 845 Key words: phosphodiesterases; DARPP-32; dopamine- GluR1 after dopamine D1 receptor agonist or forskolin stimu- stimulated locomotor activity; spatial learning and memory; lation. PDE1B Ϫ/Ϫ and PDE1B ϩ/Ϫ mice demonstrated Morris Morris water maze; methamphetamine; SKF81297; forskolin Calcium/calmodulin-dependent phosphodiesterases (CaM- (CaMKII) and calcineurin and have the potential to activate PDEs) are members of one of 11 families of PDEs (Soderling et CaM-PDEs. Dopamine D1 or D2 receptor activation leads to al., 1999;Yuasa et al., 2001) and comprise the only family that acts adenylyl cyclase activation or inhibition, respectively (Traficante ϩ as a potential point of interaction between the Ca 2 and cyclic et al., 1976; Monsma et al., 1990; Cunningham and Kelley, 1993; nucleotide signaling pathways. -
PDE4A, Active Human Recombinant Protein Expressed in Sf9 Cells
Catalog # Aliquot Size P92-31G -05 5 µg P92-31G -10 10 µg PDE4A, Active Human recombinant protein expressed in Sf9 cells Catalog # P92-31G Lot # E3321-2 Product Description Specific Activity Recombinant human PDE4A (332-end) was expressed by 1,400,000 baculovirus in Sf9 insect cells using an N-terminal GST tag. The gene accession number is NM_001111307. 1,050,000 Gene Aliases 700,000 350,000 PDE4; DPDE2; PDE46 (RLU) Activity 0 Formulation 3 4.2 5.4 6.6 7.8 9 Protein (ng) Recombinant protein stored in 50mM Tris-HCl, pH 7.5, 150mM NaCl, 10mM glutathione, 0.1mM EDTA, 0.25mM The specific activity of PDE4A was determined to be 1100 nmol DTT, 0.1mM PMSF, 25% glycerol. /min/mg as per activity assay protocol. Storage and Stability Purity o Store product at –70 C. For optimal storage, aliquot target into smaller quantities after centrifugation and store at recommended temperature. For most favorable performance, avoid repeated handling and multiple The purity was determined to be freeze/thaw cycles. >75% by densitometry. Approx. MW 110kDa. Scientific Background PDE4A is a member of the phosphodiesterase family of proteins that play a critical role in regulating intracellular levels of cAMP. In vitro phosphorylation of PDE4A by the PDE4A, Active PKA-catalytic subunit increases the enzyme's sensitivity to Human recombinant protein expressed in Sf9 cells Mg(2+), leading to a 4-fold increase in cAMP hydrolysis without affecting the Km. PDE4 is widely expressed in Catalog # P92-31G brain tumors and promotes their growth and treatment Specific Activity 1100 nmol/min/mg with PDE4A inhibitor Rolipram overcomes tumor resistance and mediates tumor regression (1). -
Targets and Mechanisms Validated Trials on the Horizon Targets And
TargetsTargets and and Mechanisms Mechanisms Validated Validated TrialsTrials on on the the Horizon Horizon Research Investors Report 2011 TABLE OF CONTENTS Huntington’s Disease Research in 2011: Targets and Mechanisms Validated; Trials on the Horizon FINDING AND VALIDATING TARGETS 4 DRP1 4 Ku705 5 A CLOSER LOOK AT THE HD PROTEIN 6 HR Protein aggregates visualized 6 Form of the HD protein associated with neurodegeneration identified 7 DISCOVERING/DEVELOPING NEW DRUGS AND UNDERSTANDING THEIR MECHANISMS 10 Dantrolene appears to be neuroprotective 10 Melatonin delays onset and prolongs survival in the R6/2 Mouse 11 KMO Inhibitor developed 12 New Caspase Inhibitors identified and Optimized 14 Quinazoline derivative looks promising 15 Phosphodiesterase-10 inhibitors 16 Novel benzoxazine compounds may be neuroprotective 18 Dimethylfumarate is helpful to the YAC128 Mouse 18 IPSC Consortium creates stem cell lines 19 FINDING AND VALIDATING BIOMARKERS 21 H2AFY 21 Protein Aggregates 22 TRACK-HD 22 UNDERSTANDING THE DISEASE COURSE AND IMPROVING CLINICAL MEASURES 25 Progression of HD and MRI imaging 25 Enroll-HD 25 POTENTIAL TREATMENTS MOVING CLOSE TO CLINICAL TRIALS 27 RNAi Primate Study 27 ASOs can be made allele specific 27 Mesenchymal stem cells with BDNF 28 CLINICAL TRIALS 30 Lessons from Dimebon 30 Neurosearch -- Huntexil (ACR-16) 31 Prana Biotech Copper Chelator 31 Siena Biotech Sirt1 Inhibitor 32 THE OTHER MEMBERS OF THE RESEARCH TEAM - THE PARTICIPANTS 33 FINAL THOUGHTS 35 HD DRUG DEVELOPMENT PIPELINE CHART 36 HDSA COALITION FOR THE CURE 37 HDSA CENTERS OF EXCELLENCE 38 Cover Photo: Polarizing optical microscopy image of huntingtin peptide aggregates stained with Congo Red. Courtesy of Dr. -
6-Methoxypyrrolidinylquinazoline for Imaging of Phosphodiesterase 10A with PET
Pharmaceuticals 2012, 5, 169-188; doi:10.3390/ph5020169 OPEN ACCESS Pharmaceuticals ISSN 1424-8247 www.mdpi.com/journal/pharmaceuticals Article Radiosynthesis and Radiotracer Properties of a 7-(2-[18F]Fluoroethoxy)-6-methoxypyrrolidinylquinazoline for Imaging of Phosphodiesterase 10A with PET Uta Funke 1,*, Winnie Deuther-Conrad 1, Gregor Schwan 2, Aurélie Maisonial 1, Matthias Scheunemann 1, Steffen Fischer 1, Achim Hiller 1, Detlef Briel 2 and Peter Brust 1 1 Institute of Radiopharmacy, Research Site Leipzig, Helmholtz-Zentrum Dresden-Rossendorf, Permoserstraße 15, Leipzig 04318, Germany; E-Mails: [email protected] (W.D.-C.); [email protected] (A.M.); [email protected] (M.S.); [email protected] (S.F.); [email protected] (A.H.); [email protected] (P.B.) 2 Institute of Pharmacy, Universität Leipzig, Brüderstraße 34, Leipzig 04103, Germany; E-Mails: [email protected] (G.S.); [email protected] (D.B.) * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +49-341-235-3695; Fax: +49-341-235-2731. Received: 24 November 2011; in revised form: 18 January 2012 / Accepted: 19 January 2012 / Published: 6 February 2012 Abstract: Phosphodiesterase 10A (PDE10A) is a key enzyme of intracellular signal transduction which is involved in the regulation of neurotransmission. The molecular imaging of PDE10A by PET is expected to allow a better understanding of physiological and pathological processes related to PDE10A expression and function in the brain. The aim of this study was to develop a new 18F-labeled PDE10A ligand based on a 6,7-dimethoxy- 4-pyrrolidinylquinazoline and to evaluate its properties in biodistribution studies. -
Analyses of PDE-Regulated Phosphoproteomes Reveal Unique and Specific Camp-Signaling Modules in T Cells
Analyses of PDE-regulated phosphoproteomes reveal unique and specific cAMP-signaling modules in T cells Michael-Claude G. Beltejara, Ho-Tak Laua, Martin G. Golkowskia, Shao-En Onga, and Joseph A. Beavoa,1 aDepartment of Pharmacology, University of Washington, Seattle, WA 98195 Contributed by Joseph A. Beavo, May 28, 2017 (sent for review March 10, 2017; reviewed by Paul M. Epstein, Donald H. Maurice, and Kjetil Tasken) Specific functions for different cyclic nucleotide phosphodiester- to bias T-helper polarization toward Th2, Treg, or Th17 pheno- ases (PDEs) have not yet been identified in most cell types. types (13, 14). In a few cases increased cAMP may even potentiate Conventional approaches to study PDE function typically rely on the T-cell activation signal (15), particularly at early stages of measurements of global cAMP, general increases in cAMP- activation. Recent MS-based proteomic studies have been useful dependent protein kinase (PKA), or the activity of exchange in characterizing changes in the phosphoproteome of T cells under protein activated by cAMP (EPAC). Although newer approaches various stimuli such as T-cell receptor stimulation (16), prosta- using subcellularly targeted FRET reporter sensors have helped glandin signaling (17), and oxidative stress (18), so much of the define more compartmentalized regulation of cAMP, PKA, and total Jurkat phosphoproteome is known. Until now, however, no EPAC, they have limited ability to link this regulation to down- information on the regulation of phosphopeptides by PDEs has stream effector molecules and biological functions. To address this been available in these cells. problem, we have begun to use an unbiased mass spectrometry- Inhibitors of cAMP PDEs are useful tools to study PKA/EPAC- based approach coupled with treatment using PDE isozyme- mediated signaling, and selective inhibitors for each of the 11 PDE – selective inhibitors to characterize the phosphoproteomes of the families have been developed (19 21). -
The Rational Search for PDE10A Inhibitors from Sophora Flavescens Roots Using Pharmacophore‑ and Docking‑Based Virtual Screening
388 MOLECULAR MEDICINE REPORTS 17: 388-393, 2018 The rational search for PDE10A inhibitors from Sophora flavescens roots using pharmacophore‑ and docking‑based virtual screening HAN-TIAN FAN1, JUN-FANG GUO1, YU-XIN ZHANG2, YU-XI GU1, ZHONG-QI NING1, YAN-JIANG QIAO2 and XING WANG1,3 1School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069; 2Key Laboratory of TCM-Information Engineer of State Administration of TCM, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100102; 3Beijing Key Laboratory of Traditional Chinese Medicine (TCM) Collateral Disease Theory Research, Capital Medical University, Beijing 100069, P.R. China Received May 27, 2017; Accepted August 31, 2017 DOI: 10.3892/mmr.2017.7871 Abstract. Phosphodiesterase 10A (PDE10A) has been Introduction confirmed to be an important target for the treatment of central nervous system (CNS) disorders. The purpose of the Phosphodiesterases (PDEs) are a family of enzymes that are present study was to identify PDE10A inhibitors from herbs able to lyse phosphodiester bonds, are expressed widely, and used in traditional Chinese medicine. Pharmacophore and have demonstrable clinical significance (1,2). PDE10A has molecular docking techniques were used to virtually screen been demonstrated to be a potential target for the treatment the chemical molecule database of Sophora flavescens, a of central nervous system (CNS) disorders (3,4). Previous well-known Chinese herb that has been used for improving studies have confirmed that PDE10A inhibitors have impor- mental health and regulating the CNS. The pharmacophore tant biological activity in the treatment of psychosis (5-7). model generated recognized the common functional groups of Therefore, screening for PDE10A inhibitors is an effective known PDE10A inhibitors. -
Peripheral Blood Gene Expression Patterns Discriminate Among
Mesko et al. BMC Medical Genomics 2010, 3:15 http://www.biomedcentral.com/1755-8794/3/15 RESEARCH ARTICLE Open Access PeripheralResearch article blood gene expression patterns discriminate among chronic inflammatory diseases and healthy controls and identify novel targets Bertalan Mesko†1, Szilard Poliska1†1,4, Andrea Szegedi3, Zoltan Szekanecz5, Karoly Palatka6, Maria Papp6 and Laszlo Nagy*1,2,4 Abstract Background: Chronic inflammatory diseases including inflammatory bowel disease (IBD; Crohn's disease and ulcerative colitis), psoriasis and rheumatoid arthritis (RA) afflict millions of people worldwide, but their pathogenesis is still not well understood. It is also not well known if distinct changes in gene expression characterize these diseases and if these patterns can discriminate between diseased and control patients and/or stratify the disease. The main focus of our work was the identification of novel markers that overlap among the 3 diseases or discriminate them from each other. Methods: Diseased (n = 13, n = 15 and n = 12 in IBD, psoriasis and RA respectively) and healthy patients (n = 18) were recruited based on strict inclusion and exclusion criteria; peripheral blood samples were collected by clinicians (30 ml) in Venous Blood Vacuum Collection Tubes containing EDTA and peripheral blood mononuclear cells were separated by Ficoll gradient centrifugation. RNA was extracted using Trizol reagent. Gene expression data was obtained using TaqMan Low Density Array (TLDA) containing 96 genes that were selected by an algorithm and the statistical analyses were performed in Prism by using non-parametric Mann-Whitney U test (P-values < 0.05). Results: Here we show that using a panel of 96 disease associated genes and measuring mRNA expression levels in peripheral blood derived mononuclear cells; we could identify disease-specific gene panels that separate each disease from healthy controls. -
Phosphodiesterase (PDE)
Phosphodiesterase (PDE) Phosphodiesterase (PDE) is any enzyme that breaks a phosphodiester bond. Usually, people speaking of phosphodiesterase are referring to cyclic nucleotide phosphodiesterases, which have great clinical significance and are described below. However, there are many other families of phosphodiesterases, including phospholipases C and D, autotaxin, sphingomyelin phosphodiesterase, DNases, RNases, and restriction endonucleases, as well as numerous less-well-characterized small-molecule phosphodiesterases. The cyclic nucleotide phosphodiesterases comprise a group of enzymes that degrade the phosphodiester bond in the second messenger molecules cAMP and cGMP. They regulate the localization, duration, and amplitude of cyclic nucleotide signaling within subcellular domains. PDEs are therefore important regulators ofsignal transduction mediated by these second messenger molecules. www.MedChemExpress.com 1 Phosphodiesterase (PDE) Inhibitors, Activators & Modulators (+)-Medioresinol Di-O-β-D-glucopyranoside (R)-(-)-Rolipram Cat. No.: HY-N8209 ((R)-Rolipram; (-)-Rolipram) Cat. No.: HY-16900A (+)-Medioresinol Di-O-β-D-glucopyranoside is a (R)-(-)-Rolipram is the R-enantiomer of Rolipram. lignan glucoside with strong inhibitory activity Rolipram is a selective inhibitor of of 3', 5'-cyclic monophosphate (cyclic AMP) phosphodiesterases PDE4 with IC50 of 3 nM, 130 nM phosphodiesterase. and 240 nM for PDE4A, PDE4B, and PDE4D, respectively. Purity: >98% Purity: 99.91% Clinical Data: No Development Reported Clinical Data: No Development Reported Size: 1 mg, 5 mg Size: 10 mM × 1 mL, 10 mg, 50 mg (R)-DNMDP (S)-(+)-Rolipram Cat. No.: HY-122751 ((+)-Rolipram; (S)-Rolipram) Cat. No.: HY-B0392 (R)-DNMDP is a potent and selective cancer cell (S)-(+)-Rolipram ((+)-Rolipram) is a cyclic cytotoxic agent. (R)-DNMDP, the R-form of DNMDP, AMP(cAMP)-specific phosphodiesterase (PDE) binds PDE3A directly.