DOCSLIB.ORG

Comprehensive GPCR Assay Solutions for Drug Discovery

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Comprehensive GPCR Assay Solutions for Drug Discovery GPCR Screening with Confidence Comprehensive GPCR Assay Solutions for Drug Discovery Why go anywhere else? Why go anywhere else? We cover the entire range of GPCR assay possibilities. Shop at the GPCR Super Store. From PerkinElmer — your one stop solution provider. PerkinElmer — a Source of Confidence for Your GPCR Research and Discovery G-Protein Coupled Receptors (GPCRs) continue to be significant therapeutic Contents targets, and thus GPCR assays and screening campaigns remain major •Assay Platforms for GPCR Binding Assays ...........2 components of drug discovery research • NEN Radioligands for GPCR Binding Assays .......4 programs worldwide. The need for • Cloned Receptors for GPCR Binding Assays.........4 reliable, proven, and versatile assay • Adenylyl Cyclase Activation/Inhibition and technologies and reagents is a key cAMP Quantitation ................................................5 component for successful and productive •Detection Instrumentation to Measure research. That’s why PerkinElmer GPCR Assays ........................................................11 has assembled the world’s most •Ordering Information: comprehensive product offering for DELFIA Ligands and Reagents.............................16 GPCR research and drug discovery, covering the entire range of GPCR GPCR-Specific NEN Radioligands.......................17 assay possibilities. SignalScreen® Cloned Receptors .........................18 Membrane Target Systems Cloned Receptors .....23 Our products and expertise span receptor ligand FlashBlue™ Scintillating Beads and binding assays through functional cellular assays. FlashPlate Microplates.........................................27 Our range of assay platforms and detection technologies Adenylyl Cyclase Assay Systems for includes essential radioisotopic technologies and state- Cell-Based Functional cAMP Measurement.........28 of-the-art luminescence and fluorescence alternatives. cAMP Assay Systems for cAMP Quantitation These are supported by a comprehensive range of in Plasma, Urine and Cell Culture Samples .........28 reagents including NEN® Radioligands, DELFIA® GTP Binding Assay Systems................................28 ligands and cloned receptor membranes, assay systems IP Functional Assay System ...............................29 using our AlphaScreen™, LANCE™, DELFIA, FlashPlate® 3 2 Reporter Gene Luminescence Assay Systems ......29 and [FP]™ technologies, plus versatile, scalable detec- tion instrumentation. Combined with our demonstrated Microplates for Filtration Assays and dedication to new product introductions— ligands, Cell-Based Assays ................................................29 receptors, functional assays, and technologies— you can be confident we’ll always be there to support this essential research area. You won’t find this commitment to GPCR research anywhere else. With PerkinElmer behind your GPCR assays, you can screen with confidence, today and in the future. Why go anywhere else? www.perkinelmer.com 1 Your Choice of Assay Platforms for GPCR Binding Assays PerkinElmer supports the widest range of assay tech- And, we’re the first to offer a viable and robust alterna- niques for GPCR binding assays. Our scintillation tive to radioisotopic assays, using our DELFIA technology, proximity assay platforms include FlashBlue™ beads a heterogeneous time resolved fluorescence assay and FlashPlate and Image FlashPlate microplates. platform. All of these options can now supplement These are all compatible with our extensive line of traditional filtration assays to increase the performance NEN Radioligands and SignalScreen® and Membrane and productivity of your GPCR assays. Target Systems (MTS) cloned receptor preparations. PerkinElmer Assay Platforms for Receptor Ligand Binding Assays Selection Criteria DELFIA FlashBlue FlashPlate Image Filtration Technology Beads Microplates FlashPlate Assays Microplates Homogeneous No Yes Yes Yes No Non-Rad Yes No No No No Format 96- and 384- 96- and 384- 96- and 384- 384-shallow 96-well plates well plates well plates well plates well plates Throughput >40,000 >20,000 >20,000 >50,000 >15,000 (wells in an 8-hour day) Recommended DELFIA Clear Bottom or N/A N/A UniFilter® Microplate Type AcroWell™ White OptiPlate Filter Plates Filter Plates Microplates Stability of Signal >24 hours >24 hours >24 hours >24 hours >24 hours DELFIA® TRF Technology Our DELFIA technology is a robust, time-resolved, Systems (MTS) cloned receptors. For easy assay setup, fluorometric assay (TRF) technology based on dissociative use DELFIA L*R Wash Solution and DELFIA L*R fluorescence enhancement using lanthanide chelates. Binding Buffer. After incubation unbound ligand is Invented by and available exclusively from PerkinElmer, removed by filtration, then signal is enhanced using it’s been widely adopted for many HTS applications DELFIA Enhancement Solution. It’s simple and reliable. due to its extremely high sensitivity and wide dynamic range. Receptor binding assays using DELFIA technology have been proven to be a robust non-radioactive alterna- tive to traditional receptor-radioligand binding assays. Europium-labeled ligands are stable reagents for DELFIA receptor-ligand binding assays. The binding assay may be done in an AcroWell FilterPlate using PerkinElmer’s SignalScreen or Membrane Target 2 FlashBlue™ GPCR Scintillating Beads Our FlashBlue GPCR beads are scintillating beads binding assays, radioligand molecules bound to the specifically designed and proven for high throughput membranes’ receptors activate the scintillating beads homogeneous GPCR-radioligand binding assays. Three and trigger a blue light emission. Assays can be read microns in diameter, they consist of a polystyrene core on PerkinElmer’s TopCount® and MicroBeta® Liquid covered by a hydrophilic coating that minimizes non- Scintillation and Luminescence Counters in 96- and specific binding. Wheat germ agglutinin (WGA) is 384-well formats, as well as our TriCarb LSC for covalently attached to the surface of FlashBlue GPCR tube-based assays. beads for the capture of cellular membranes. In GPCR FlashPlate® and Image FlashPlate® Microplates FlashPlate Microplates 125 3 35 33 14 Our popular FlashPlate microplates are designed for of a variety of isotopes (e.g., I, H, S, P and C) high-volume, homogeneous radioligand binding assays, without the addition of liquid scintillation cocktail. based on the principle of scintillation proximity. The FlashPlate microplates can be read on TopCount or interior of each well is permanently coated with a MicroBeta counters. thin layer of plastic scintillant, enabling detection Image FlashPlate Microplates Our 384-well Image FlashPlate microplate is coated The FlashPlate platform is especially suitable for with a special scintillant that emits in the red region of receptor binding assays using highly purified membrane the spectrum. This characteristic allows detection with preparations and/or receptor preparations known imaging readers, such as the PerkinElmer ViewLux™ to contain moderate to higher concentrations of ultraHTS Microplate Imager, enabling very high testing receptor protein. Use of FlashPlate pre-coated with throughput. Image FlashPlate also eliminates or greatly wheat germ agglutinin allows greater flexibility in reduces interference from colored compounds. These receptor choice and optimal ease-of-use. plates have shallower wells to permit lower volume assays and, therefore, use less reagent, which is ideal for further miniaturization. ScreenReady Targets™ ScreenReady Targets™ provide optimum performance of recombinant GPCR membranes pre-coated onto with FlashPlate microplates. They’re ready-to-use, fully FlashPlate microplates, the appropriate radioligand, 125 validated GPCR receptor binding assays that employ I and a validated assay protocol. For more information, 3 and H radioligands. This easy-to-use platform consists please contact [email protected]. www.perkinelmer.com 3 NEN Radioligands Keep Pace with Drug Discovery From the company that pioneered development work with you, scientist to scientist, to prepare a of the essential tools for receptor ligand binding, custom radioligand that meets your specifications. 3 125 PerkinElmer now offers over 400 NEN H and I A confidentiality agreement is available if required. ligands. A continuous pipeline of new product We are committed to your success. introductions ensures we’ll always keep pace with Please refer to the tables on pages 17–18 for a listing of discoveries and trends in receptor research and our available GPCR membranes with their compatible drug discovery. Each of our ultrapure radioligands NEN Radioligands. PerkinElmer also offers a number is fully characterized for pharmacological action of radioligands, some exclusive, specific for GPCRs and validated in a receptor binding assay. that are not found in the aforementioned tables. For If you don’t find exactly what you need, we invite our complete radioligand offering, please visit our you to take advantage of our custom services. We’ll website at www.perkinelmer.com/nenradioligands. World’s Most Comprehensive Line of Cloned Receptors for GPCR Binding Assays PerkinElmer offers more than 170 cloned receptor They provide excellent targets for GPCR assays using membrane preparations, with a constant flow of new the technique of your choice, including: receptors as they are identified. Our line includes • Receptor radioligand binding assays with FlashPlate over 150 GPCRs as well as several tyrosine kinase, microplates, FlashBlue beads,
Load more

Recommended publications
  • TABLE 1 Studies of Antagonist Activity in Constitutively Active
    TABLE 1 Studies of antagonist activity in constitutively active receptors systems shown to demonstrate inverse agonism for at least one ligand Targets are natural Gs and constitutively active mutants (CAM) of GPCRs. Of 380 antagonists, 85% of the ligands demonstrate inverse agonism. Receptor Neutral Antagonist Inverse Agonist Reference Human β2-adrenergic Dichloroisoproterenol, pindolol, labetolol, timolol, Chidiac et al., 1996; Azzi et alprenolol, propranolol, ICI 118,551, cyanopindolol al., 2001 Turkey erythrocyte β-adrenergic Propranolol, pindolol Gotze et al., 1994 Human β2-adrenergic (CAM) Propranolol Betaxolol, ICI 118,551, sotalol, timolol Samama et al., 1994; Stevens and Milligan, 1998 Human/guinea pig β1-adrenergic Atenolol, propranolol Mewes et al., 1993 Human β1-adrenergic Carvedilol CGP20712A, metoprolol, bisoprolol Engelhardt et al., 2001 Rat α2D-adrenergic Rauwolscine, yohimbine, WB 4101, idazoxan, Tian et al., 1994 phentolamine, Human α2A-adrenergic Napthazoline, Rauwolscine, idazoxan, altipamezole, levomedetomidine, Jansson et al., 1998; Pauwels MPV-2088 (–)RX811059, RX 831003 et al., 2002 Human α2C-adrenergic RX821002, yohimbine Cayla et al., 1999 Human α2D-adrenergic Prazosin McCune et al., 2000 Rat α2-adrenoceptor MK912 RX821002 Murrin et al., 2000 Porcine α2A adrenoceptor (CAM- Idazoxan Rauwolscine, yohimbine, RX821002, MK912, Wade et al., 2001 T373K) phentolamine Human α2A-adrenoceptor (CAM) Dexefaroxan, (+)RX811059, (–)RX811059, RS15385, yohimbine, Pauwels et al., 2000 atipamezole fluparoxan, WB 4101 Hamster α1B-adrenergic
    [Show full text]
  • Subanesthetic Doses of Ketamine Transiently Decrease Serotonin Transporter Activity: a PET Study in Conscious Monkeys
    Neuropsychopharmacology (2013) 38, 2666–2674 & 2013 American College of Neuropsychopharmacology. All rights reserved 0893-133X/13 www.neuropsychopharmacology.org Subanesthetic Doses of Ketamine Transiently Decrease Serotonin Transporter Activity: A PET Study in Conscious Monkeys 1 1 1 1 1 Shigeyuki Yamamoto , Hiroyuki Ohba , Shingo Nishiyama , Norihiro Harada , Takeharu Kakiuchi , 1 ,2 Hideo Tsukada and Edward F Domino* 1 2 Central Research Laboratory, Hamamatsu Photonics KK, Hamakita, Japan; Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA Subanesthetic doses of ketamine, an N-methyl-D-aspartic acid (NMDA) antagonist, have a rapid antidepressant effect which lasts for up to 2 weeks. However, the neurobiological mechanism regarding this effect remains unclear. In the present study, the effects of subanesthetic doses of ketamine on serotonergic systems in conscious monkey brain were investigated. Five young monkeys 11 underwent four positron emission tomography measurements with [ C]-3-amino-4-(2-dimethylaminomethyl-phenylsulfanyl)benzoni- 11 trile ([ C]DASB) for the serotonin transporter (SERT), during and after intravenous infusion of vehicle or ketamine hydrochloride in a 11 dose of 0.5 or 1.5 mg/kg for 40 min, and 24 h post infusion. Global reduction of [ C]DASB binding to SERT was observed during ketamine infusion in a dose-dependent manner, but not 24 h later. The effect of ketamine on the serotonin 1A receptor (5-HT1A-R) and dopamine transporter (DAT) was also investigated in the same subjects studied with [11C]DASB. No significant changes were observed in either 5-HT -R or DAT binding after ketamine infusion. Microdialysis analysis indicated that ketamine infusion transiently increased 1A serotonin levels in the extracellular fluid of the prefrontal cortex.
    [Show full text]
  • Zebrafish Behavioral Profiling Links Drugs to Biological Targets and Rest/Wake Regulation
    www.sciencemag.org/cgi/content/full/327/5963/348/DC1 Supporting Online Material for Zebrafish Behavioral Profiling Links Drugs to Biological Targets and Rest/Wake Regulation Jason Rihel,* David A. Prober, Anthony Arvanites, Kelvin Lam, Steven Zimmerman, Sumin Jang, Stephen J. Haggarty, David Kokel, Lee L. Rubin, Randall T. Peterson, Alexander F. Schier* *To whom correspondence should be addressed. E-mail: [email protected] (A.F.S.); [email protected] (J.R.) Published 15 January 2010, Science 327, 348 (2010) DOI: 10.1126/science.1183090 This PDF file includes: Materials and Methods SOM Text Figs. S1 to S18 Table S1 References Supporting Online Material Table of Contents Materials and Methods, pages 2-4 Supplemental Text 1-7, pages 5-10 Text 1. Psychotropic Drug Discovery, page 5 Text 2. Dose, pages 5-6 Text 3. Therapeutic Classes of Drugs Induce Correlated Behaviors, page 6 Text 4. Polypharmacology, pages 6-7 Text 5. Pharmacological Conservation, pages 7-9 Text 6. Non-overlapping Regulation of Rest/Wake States, page 9 Text 7. High Throughput Behavioral Screening in Practice, page 10 Supplemental Figure Legends, pages 11-14 Figure S1. Expanded hierarchical clustering analysis, pages 15-18 Figure S2. Hierarchical and k-means clustering yield similar cluster architectures, page 19 Figure S3. Expanded k-means clustergram, pages 20-23 Figure S4. Behavioral fingerprints are stable across a range of doses, page 24 Figure S5. Compounds that share biological targets have highly correlated behavioral fingerprints, page 25 Figure S6. Examples of compounds that share biological targets and/or structural similarity that give similar behavioral profiles, page 26 Figure S7.
    [Show full text]
  • The Role of the Serotonergic System and the Effects of Antidepressants During Brain Development Examined Using in Vivo Pet Imaging and in Vitro Receptor Binding
    From THE DEPARTMENT OF CLINICAL NEUROSCIENCE Karolinska Institutet, Stockholm, Sweden THE ROLE OF THE SEROTONERGIC SYSTEM AND THE EFFECTS OF ANTIDEPRESSANTS DURING BRAIN DEVELOPMENT EXAMINED USING IN VIVO PET IMAGING AND IN VITRO RECEPTOR BINDING Stal Saurav Shrestha Stockholm 2014 Cover Illustration: Voxel-wise analysis of the whole monkey brain using the PET radioligand, [11C]DASB showing persistent serotonin transporter upregulation even after more than 1.5 years of fluoxetine discontinuation. All previously published papers were reproduced with permission from the publisher. Published by Karolinska Institutet. Printed by Universitetsservice-AB © Stal Saurav Shrestha, 2014 ISBN 978-91-7549-522-4 Serotonergic System and Antidepressants During Brain Development To my family Amaze yourself ! Stal Saurav Shrestha, 2014 The Department of Clinical Neuroscience The role of the serotonergic system and the effects of antidepressants during brain development examined using in vivo PET imaging and in vitro receptor binding AKADEMISK AVHANDLING som för avläggande av medicine doktorsexamen vid Karolinska Institutet offentligen försvaras i CMM föreläsningssalen L8:00, Karolinska Universitetssjukhuset, Solna THESIS FOR DOCTORAL DEGREE (PhD) Stal Saurav Shrestha Date: March 31, 2014 (Monday); Time: 10 AM Venue: Center for Molecular Medicine Lecture Hall Floor 1, Karolinska Hospital, Solna Principal Supervisor: Opponent: Robert B. Innis, MD, PhD Klaus-Peter Lesch, MD, PhD National Institutes of Health University of Würzburg Department of NIMH Department
    [Show full text]
  • Properties of Chemically Oxidized Kininogens*
    Vol. 50 No. 3/2003 753–763 QUARTERLY Properties of chemically oxidized kininogens*. Magdalena Nizio³ek, Marcin Kot, Krzysztof Pyka, Pawe³ Mak and Andrzej Kozik½ Faculty of Biotechnology, Jagiellonian University, Kraków, Poland Received: 30 May, 2003; revised: 01 August, 2003; accepted: 11 August, 2003 Key words: bradykinin, N-chlorosuccinimide, chloramine-T, kallidin, kallikrein, reactive oxygen species Kininogens are multifunctional proteins involved in a variety of regulatory pro- cesses including the kinin-formation cascade, blood coagulation, fibrynolysis, inhibi- tion of cysteine proteinases etc. A working hypothesis of this work was that the prop- erties of kininogens may be altered by oxidation of their methionine residues by reac- tive oxygen species that are released at the inflammatory foci during phagocytosis of pathogen particles by recruited neutrophil cells. Two methionine-specific oxidizing reagents, N-chlorosuccinimide (NCS) and chloramine-T (CT), were used to oxidize the high molecular mass (HK) and low molecular mass (LK) forms of human kininogen. A nearly complete conversion of methionine residues to methionine sulfoxide residues in the modified proteins was determined by amino acid analysis. Production of kinins from oxidized kininogens by plasma and tissue kallikreins was significantly lower (by at least 70%) than that from native kininogens. This quenching effect on kinin release could primarily be assigned to the modification of the critical Met-361 residue adja- cent to the internal kinin sequence in kininogen. However, virtually no kinin could be formed by human plasma kallikrein from NCS-modified HK. This observation sug- gests involvement of other structural effects detrimental for kinin production. In- deed, NCS-oxidized HK was unable to bind (pre)kallikrein, probably due to the modifi- cation of methionine and/or tryptophan residues at the region on the kininogen mole- cule responsible for the (pro)enzyme binding.
    [Show full text]
  • A Nonlinear Relationship Between Cerebral Serotonin Transporter And
    The Journal of Neuroscience, March 3, 2010 • 30(9):3391–3397 • 3391 Cellular/Molecular A Nonlinear Relationship between Cerebral Serotonin Transporter and 5-HT2A Receptor Binding: An In Vivo Molecular Imaging Study in Humans David Erritzoe,1,3 Klaus Holst,3,4 Vibe G. Frokjaer,1,3 Cecilie L. Licht,1,3 Jan Kalbitzer,1,3 Finn Å. Nielsen,3,5 Claus Svarer,1,3 Jacob Madsen,2 and Gitte M. Knudsen1,3 1Neurobiology Research Unit and 2PET and Cyclotron Unit, University Hospital Rigshospitalet, DK-2100 Copenhagen, Denmark, 3Center for Integrated Molecular Brain Imaging, DK-2100 Copenhagen, Denmark, 4Department of Biostatistics, University of Copenhagen, DK-2200 Copenhagen, Denmark, and 5DTU Informatics, Technical University of Denmark, DK-2800 Lyngby, Denmark Serotonergic neurotransmission is involved in the regulation of physiological functions such as mood, sleep, memory, and appetite. Withintheserotonintransmittersystem,boththepostsynapticallylocatedserotonin2A(5-HT2A )receptorandthepresynapticserotonin transporter (SERT) are sensitive to chronic changes in cerebral 5-HT levels. Additionally, experimental studies suggest that alterations in either the 5-HT2A receptor or SERT level can affect the protein level of the counterpart. The aim of this study was to explore the covariation betweencerebral5-HT2A receptorandSERT invivointhesamehealthyhumansubjects.Fifty-sixhealthyhumansubjectswithameanage of 36 Ϯ 19 years were investigated. The SERT binding was imaged with [ 11C]3-amino-4-(2-dimethylaminomethyl-phenylsulfanyl)- 18 benzonitrile (DASB) and 5-HT2A receptor binding with [ F]altanserin using positron emission tomography. Within each individual, a regionalintercorrelationforthevariousbrainregionswasseenwithbothmarkers,mostnotablyfor5-HT2A receptorbinding.Aninverted U-shaped relationship between the 5-HT2A receptor and the SERT binding was identified. The observed regional intercorrelation for both the 5-HT2A receptor and the SERT cerebral binding suggests that, within the single individual, each marker has a set point adjusted through a common regulator.
    [Show full text]
  • Sized Neuropeptides
    M ETHODS IN MOLECULAR BIOLOGY™ Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK For further volumes: http://www.springer.com/series/7651 Neuropeptides Methods and Protocols Edited by Adalberto Merighi Dipartimento di Morfofisiologia Veterinaria, Università degli Studi di Torino, Grugliasco, TO, Italy; Istituto Nazionale di Neuroscienze (INN), Università degli Studi di Torino, Grugliasco, TO, Italy Editor Adalberto Merighi Dipartimento di Morfofisiologia Veterinaria Università degli Studi di Torino and Istituto Nazionale di Neuroscienze (INN) Università degli Studi di Torino Grugliasco, TO, Italy [email protected] Please note that additional material for this book can be downloaded from http://extras.springer.com ISSN 1064-3745 e-ISSN 1940-6029 ISBN 978-1-61779-309-7 e-ISBN 978-1-61779-310-3 DOI 10.1007/978-1-61779-310-3 Springer New York Dordrecht Heidelberg London Library of Congress Control Number: 2011936011 © Springer Science+Business Media, LLC 2011 All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights.
    [Show full text]
  • Thesis (1.852Mb)
    From DEPARTMENT OF CLINICAL NEUROSCIENCE Karolinska Institutet, Stockholm, Sweden PET STUDIES ON THE MECHANISMS OF ACTION OF ANTIDEPRESSANT AND ANTIPSYCHOTIC DRUGS Kai-Chun Yang Stockholm 2017 Cover illusion: Parasagittal PET-section of a nonhuman primate brain showing regional reductions of 11 [ C]AZ10419369 binding to the 5-HT1B receptor after administration of vortioxetine, a novel antidepressant. All previously published papers were reproduced with permission from the publisher. Published by Karolinska Institutet. Printed by E-Print AB 2017 © Kai-Chun Yang, 2017 ISBN 978-91-7676-786-3 PET Studies on the Mechanisms of Action of Antidepressant and Antipsychotic Drugs THESIS FOR DOCTORAL DEGREE (Ph.D.) By Kai-Chun Yang Principal Supervisor: Opponent: Dr Sjoerd J. Finnema Professor Yasuyoshi Watanabe Karolinska Institutet RIKEN Department of Clinical Neuroscience Center for Life Science Technologies Co-supervisor(s): Examination Board: Professor Lars Farde Professor Sven Ove Ögren Karolinska Institutet Karolinska Institutet Department of Clinical Neuroscience Department of Neuroscience Professor Christer Halldin Professor Hans Ågren Karolinska Institutet University of Gothenburg Department of Clinical Neuroscience Institute of Neuroscience and Physiology Dr Akihiro Takano Professor Mark Lubberink Karolinska Institutet Uppsala University Department of Clinical Neuroscience Department of Surgical Sciences In memory of my father, 楊正宗 (Yang, Cheng-Tsung), 1945-2011 “I couldn't reduce it to the freshman level. That means we don't really understand it.” Richard Feynman, 1918-1988 ABSTRACT Positron emission tomography (PET) is a non-invasive molecular imaging technique suitable for examination of neurochemical biomarkers in the living brain. Among these applications, PET studies are used to facilitate the development of novel psychotropic drugs.
    [Show full text]
  • G Protein-Coupled Receptors
    S.P.H. Alexander et al. The Concise Guide to PHARMACOLOGY 2015/16: G protein-coupled receptors. British Journal of Pharmacology (2015) 172, 5744–5869 THE CONCISE GUIDE TO PHARMACOLOGY 2015/16: G protein-coupled receptors Stephen PH Alexander1, Anthony P Davenport2, Eamonn Kelly3, Neil Marrion3, John A Peters4, Helen E Benson5, Elena Faccenda5, Adam J Pawson5, Joanna L Sharman5, Christopher Southan5, Jamie A Davies5 and CGTP Collaborators 1School of Biomedical Sciences, University of Nottingham Medical School, Nottingham, NG7 2UH, UK, 2Clinical Pharmacology Unit, University of Cambridge, Cambridge, CB2 0QQ, UK, 3School of Physiology and Pharmacology, University of Bristol, Bristol, BS8 1TD, UK, 4Neuroscience Division, Medical Education Institute, Ninewells Hospital and Medical School, University of Dundee, Dundee, DD1 9SY, UK, 5Centre for Integrative Physiology, University of Edinburgh, Edinburgh, EH8 9XD, UK Abstract The Concise Guide to PHARMACOLOGY 2015/16 provides concise overviews of the key properties of over 1750 human drug targets with their pharmacology, plus links to an open access knowledgebase of drug targets and their ligands (www.guidetopharmacology.org), which provides more detailed views of target and ligand properties. The full contents can be found at http://onlinelibrary.wiley.com/doi/ 10.1111/bph.13348/full. G protein-coupled receptors are one of the eight major pharmacological targets into which the Guide is divided, with the others being: ligand-gated ion channels, voltage-gated ion channels, other ion channels, nuclear hormone receptors, catalytic receptors, enzymes and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading.
    [Show full text]
  • The 2.1 A˚Resolution Structure of Cyanopindolol-Bound B1
    The 2.1 A˚ Resolution Structure of Cyanopindolol-Bound + b1-Adrenoceptor Identifies an Intramembrane Na Ion that Stabilises the Ligand-Free Receptor Jennifer L. Miller-Gallacher.¤a, Rony Nehme´ ., Tony Warne, Patricia C. Edwards, Gebhard F. X. Schertler¤b¤c, Andrew G. W. Leslie, Christopher G. Tate* Structural Studies Division, MRC Laboratory of Molecular Biology, Cambridge, Cambridgeshire, United Kingdom Abstract The b1-adrenoceptor (b1AR) is a G protein-coupled receptor (GPCR) that is activated by the endogenous agonists adrenaline and noradrenaline. We have determined the structure of an ultra-thermostable b1AR mutant bound to the weak partial agonist cyanopindolol to 2.1 A˚ resolution. High-quality crystals (100 mm plates) were grown in lipidic cubic phase without the assistance of a T4 lysozyme or BRIL fusion in cytoplasmic loop 3, which is commonly employed for GPCR crystallisation. An intramembrane Na+ ion was identified co-ordinated to Asp872.50, Ser1283.39 and 3 water molecules, which is part of a more extensive network of water molecules in a cavity formed between transmembrane helices 1, 2, 3, 6 and 7. Remarkably, + this water network and Na ion is highly conserved between b1AR and the adenosine A2A receptor (rmsd of 0.3 A˚), despite an overall rmsd of 2.4 A˚ for all Ca atoms and only 23% amino acid identity in the transmembrane regions. The affinity of + agonist binding and nanobody Nb80 binding to b1AR is unaffected by Na ions, but the stability of the receptor is decreased by 7.5uC in the absence of Na+. Mutation of amino acid side chains that are involved in the co-ordination of either + Na or water molecules in the network decreases the stability of b1AR by 5–10uC.
    [Show full text]
  • Adrenergic G-Protein- Coupled Receptor
    Vol 454 | 24 July 2008 | doi:10.1038/nature07101 ARTICLES Structure of a b1-adrenergic G-protein- coupled receptor Tony Warne1, Maria J. Serrano-Vega1, Jillian G. Baker2, Rouslan Moukhametzianov1, Patricia C. Edwards1, Richard Henderson1, Andrew G. W. Leslie1, Christopher G. Tate1 & Gebhard F. X. Schertler1 G-protein-coupled receptors have a major role in transmembrane signalling in most eukaryotes and many are important drug targets. Here we report the 2.7 A˚ resolution crystal structure of a b1-adrenergic receptor in complex with the high-affinity antagonist cyanopindolol. The modified turkey (Meleagris gallopavo) receptor was selected to be in its antagonist conformation and its thermostability improved by earlier limited mutagenesis. The ligand-binding pocket comprises 15 side chains from amino acid residues in 4 transmembrane a-helices and extracellular loop 2. This loop defines the entrance of the ligand-binding pocket and is stabilized by two disulphide bonds and a sodium ion. Binding of cyanopindolol to the b1-adrenergic receptor and binding of carazolol to the b2-adrenergic receptor involve similar interactions. A short well-defined helix in cytoplasmic loop 2, not observed in either rhodopsin or the b2-adrenergic receptor, directly interacts by means of a tyrosine with the highly conserved DRY motif at the end of helix 3 that is essential for receptor activation. G-protein-coupled receptors (GPCRs) are a large family of integral These structures, both containing the high affinity antagonist cara- membrane proteins that are prevalent in eukaryotes from yeast to zolol, defined the overall architecture of b2AR and the structure of the man, and function as key intermediaries in the transduction of sig- ligand-binding pocket.
    [Show full text]
  • Drugs for Primary Prevention of Atherosclerotic Cardiovascular Disease: an Overview of Systematic Reviews
    Supplementary Online Content Karmali KN, Lloyd-Jones DM, Berendsen MA, et al. Drugs for primary prevention of atherosclerotic cardiovascular disease: an overview of systematic reviews. JAMA Cardiol. Published online April 27, 2016. doi:10.1001/jamacardio.2016.0218. eAppendix 1. Search Documentation Details eAppendix 2. Background, Methods, and Results of Systematic Review of Combination Drug Therapy to Evaluate for Potential Interaction of Effects eAppendix 3. PRISMA Flow Charts for Each Drug Class and Detailed Systematic Review Characteristics and Summary of Included Systematic Reviews and Meta-analyses eAppendix 4. List of Excluded Studies and Reasons for Exclusion This supplementary material has been provided by the authors to give readers additional information about their work. © 2016 American Medical Association. All rights reserved. 1 Downloaded From: https://jamanetwork.com/ on 09/28/2021 eAppendix 1. Search Documentation Details. Database Organizing body Purpose Pros Cons Cochrane Cochrane Library in Database of all available -Curated by the Cochrane -Content is limited to Database of the United Kingdom systematic reviews and Collaboration reviews completed Systematic (UK) protocols published by by the Cochrane Reviews the Cochrane -Only systematic reviews Collaboration Collaboration and systematic review protocols Database of National Health Collection of structured -Curated by Centre for -Only provides Abstracts of Services (NHS) abstracts and Reviews and Dissemination structured abstracts Reviews of Centre for Reviews bibliographic
    [Show full text]