Non-Classical Amine Recognition Evolved in a Large Clade of Olfactory Receptors

Total Page:16

File Type:pdf, Size:1020Kb

Non-Classical Amine Recognition Evolved in a Large Clade of Olfactory Receptors 1 Non-classical amine recognition evolved in a large clade of olfactory receptors 2 3 Qian Li1, Yaw Tachie-Baffour1, Zhikai Liu1, Maude W. Baldwin2, Andrew C. Kruse3, and Stephen 4 D. Liberles1* 5 6 1Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; 7 2Department of Organismic and Evolutionary Biology, Harvard University and the Museum of 8 Comparative Zoology, Cambridge, MA 02138, USA; 9 3Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 10 Boston, MA 02115, USA 11 12 *Correspondence to [email protected], 13 phone: (617) 432-7283, fax: (617) 432-7285 14 15 Competing interests 16 The authors declare that no competing interests exist. 17 18 19 20 Biogenic amines are important signaling molecules, and the structural basis for their 21 recognition by G Protein-Coupled Receptors (GPCRs) is well understood. Amines are 22 also potent odors, with some activating olfactory trace amine-associated receptors 23 (TAARs). Here, we report that teleost TAARs evolved a new way to recognize amines in a 24 non-classical orientation. Chemical screens de-orphaned eleven zebrafish TAARs, with 25 agonists including serotonin, histamine, tryptamine, 2-phenylethylamine, putrescine, and 26 agmatine. Receptors from different clades contact ligands through aspartates on 27 transmembrane α-helices III (canonical Asp3.32) or V (non-canonical Asp5.42), and diamine 28 receptors contain both aspartates. Non-classical monoamine recognition evolved in two 29 steps: an ancestral TAAR acquired Asp5.42, gaining diamine sensitivity, and subsequently 30 lost Asp3.32. Through this transformation, the fish olfactory system dramatically 31 expanded its capacity to detect amines, ecologically significant aquatic odors. The 32 evolution of a second, alternative solution for amine detection by olfactory receptors 33 highlights the tremendous structural versatility intrinsic to GPCRs. 34 35 2 36 INTRODUCTION 37 38 Biogenic amines are key intercellular signaling molecules that regulate physiology and behavior. 39 In vertebrates, biogenic amines include adrenaline, serotonin, acetylcholine, histamine, and 40 dopamine, and these biogenic amines can be detected by G Protein-Coupled Receptors 41 (GPCRs) expressed on the plasma membrane of target cells. The biochemical and structural 42 basis for biogenic amine recognition by GPCRs is well understood and involves an essential salt 43 bridge between the ligand amino group and a highly conserved aspartic acid on the third 44 transmembrane α-helix of the receptor (Asp3.32; Ballesteros-Weinstein indexing)1,2. Mutation of 45 Asp3.32 in adrenaline, serotonin, acetylcholine, histamine, and dopamine receptors eliminates 46 amine recognition2,3, and GPCRs that recognize amines through another motif have not been 47 identified. 48 49 Trace amine-associated receptors (TAARs) are a family of vertebrate GPCRs distantly related 50 to biogenic amine receptors4. There are 15 TAARs in mouse, 6 in human, and 112 in zebrafish, 51 and in these or related species, all TAARs except TAAR1 function as olfactory receptors5-7. All 52 human TAARs (6/6), most mouse TAARs (13/15), and some zebrafish TAARs (27/112) retain 53 Asp3.32, suggesting that many TAARs would recognize amines. High throughput chemical 54 screens yielded ligands for TAAR18-10 and many olfactory TAARs that retain Asp3.32, including 55 TAAR3, TAAR4, TAAR5, TAAR7s, TAAR8s, TAAR9, and TAAR13c7,11,12. Each of these 56 receptors detects different amines, some of which evoke innate olfactory behaviors12-16. In 57 mouse, TAAR4 detects the aversive carnivore odor 2-phenylethylamine14, while TAAR5 detects 58 the attractive and sexually dimorphic mouse odor trimethylamine7,15. Knockout mice lacking 59 TAAR4 and TAAR5 lose behavioral responses to TAAR ligands identified in cell culture 60 studies13,15. 61 3 62 The selectivity of olfactory TAARs for particular amine odors suggests structural variability within 63 the ligand-binding pocket across the TAAR family. One clade of TAARs (including TAAR1, 64 TAAR3, and TAAR4) detects primary amines derived by amino acid decarboxylation, while a 65 second clade (including TAAR5, TAAR7s, TAAR8s, and TAAR9) prefers tertiary amines11. 66 Structural models and mutagenesis studies of TAAR1, TAAR7e, and TAAR7f predicted that 67 ligand binding occurs in the membrane plane, Asp3.32 forms a salt bridge to the ligand amino 68 group, and other transmembrane residues provide a scaffold that supports ligand binding and 69 enables selectivity11,17. 70 71 A third TAAR clade (clade III TAARs) arose in fish, and includes the majority (87/112) of 72 zebrafish TAARs6. Most clade III TAARs lack Asp3.32 (85/87), and it has been proposed that 73 these receptors may not detect amines6,18. However, ligands have not been identified for any of 74 these 85 receptors. More generally, ligands are known for only one zebrafish TAAR, TAAR13c, 75 which retains Asp3.32 and detects the carrion odor cadaverine12. Structure-activity analysis 76 revealed that TAAR13c preferentially recognizes odd-chained, medium-sized diamines, and 77 likely contains two distinct cation recognition sites within the ligand-binding pocket12. However, 78 the structural basis for diamine recognition by TAAR13c was unknown. 79 80 Here, we find that TAAR13c recognizes cadaverine through two remote transmembrane 81 aspartic acids, Asp3.32 and Asp5.42. A TAAR13c variant with a charge-neutralizing mutation at 82 Asp5.42 has an altered ligand preference for monoamines rather than diamines. We propose that 83 Asp3.32 and Asp5.42 together form a general di-cation recognition motif, and note this motif to be 84 present in several zebrafish, mouse, and human TAARs. Surprisingly, Asp5.42 is retained in 84 85 out of 85 zebrafish TAARs that lack Asp3.32, raising the possibility that these receptors recognize 86 amines positioned in a non-canonical inverted orientation with the amino group pointing towards 87 transmembrane α-helix V. Using a high throughput chemical screening approach, we identified 4 88 ligands for eleven additional zebrafish TAARs, and found that different receptors detect amines 89 through salt bridge contacts at Asp3.32 and/or Asp5.42. TAAR10a, TAAR10b, TAAR12h, and 90 TAAR12i are Asp3.32-containing clade I TAARs that detect serotonin, tryptamine, 2- 91 phenylethylamine, and 3-methoxytyramine, while TAAR16c, TAAR16e, and TAAR16f are 92 Asp5.42-containing clade III TAARs that detect N-methylpiperidine, N,N- 93 dimethylcyclohexylamine, and isoamylamine. TAAR13a, TAAR13d, TAAR13e, and TAAR14d 94 have both Asp3.32 and Asp5.42, and recognize the di-cationic molecules putrescine, histamine, 95 and agmatine. 96 97 Based on these findings, we propose that a large clade of olfactory GPCRs evolved a novel way 98 to detect amines that involves a non-classical salt bridge to transmembrane α-helix V. 99 Furthermore, new TAAR ligands include several biogenic amines for which other receptors were 100 identified, including serotonin, histamine, and 2-phenylethylamine. These findings illustrate that 101 different members of the GPCR superfamily can evolve divergent ligand binding pockets for 102 recognition of the same agonist. 103 104 RESULTS 105 106 Two remote amine recognition sites in the cadaverine receptor 107 108 To gain structural insights into diamine recognition by the cadaverine receptor TAAR13c, we 109 generated a TAAR13c homology model (Figure 1A) based on the X-ray crystal structure of 19 110 agonist-bound β2 adrenergic receptor (Protein Data Bank Entry 4LDE) . The β2 adrenergic 111 receptor is the closest homolog (33% amino acid identity) of TAAR13c for which an active-state 112 structure is currently available. The homology model of TAAR13c indicated a canonical fold of 5 113 seven transmembrane α-helices with ligand bound at the core. Using the homology model, we 114 performed computational docking of cadaverine to the orthosteric binding site. One cadaverine 115 amino group of the docked ligand was located near Asp1123.32, which corresponds to the highly 116 conserved amine-contact site in biogenic amine receptors. The other cadaverine amino group 117 was positioned at the opposing end of the ligand-binding pocket, 3 Å away from another 118 aspartate, Asp2025.42. The second amino group was also within 5 Å of side chains from 119 Thr2035.43 and Ser2766.55, Leu192 and Phe194 on extracellular loop 2, and the backbone of 120 Ser1995.39 (additional nearby residues are depicted in Figure 1-figure supplement 1). Based on 121 its proximity and charge, Asp2025.42 was deemed a prime candidate to function as a counterion 122 that forms a salt bridge to the second cadaverine amino group. Position 5.42 in several other 123 GPCRs directly contacts ligands through hydrogen bonds, salt bridges, or van der Waals 124 interactions3, and our homology model suggests a role for this amino acid in ligand recognition 125 by TAAR13c. 126 127 We used an established reporter gene assay7,11 to query the response properties of TAAR13c 128 variants with charge-neutralizing mutations at Asp3.32 (TAAR13cD112A) and Asp5.42 129 (TAAR13cD202A). HEK-293 cells were co-transfected with plasmids encoding TAARs and a 130 cAMP-dependent reporter gene encoding secreted alkaline phosphatase (Cre-Seap). 131 Transfected cells were exposed to test ligands, and assayed for reporter activity using a 132 fluorescent phosphatase substrate. As previously published with this paradigm12, cadaverine 133 robustly activated TAAR13c, with a half maximal response occurring at ~10 μM. In contrast, 134 cadaverine failed to activate either the TAAR13cD112A or TAAR13cD202A mutant at any 135 concentration tested, up to 1 mM (Figure 1, Figure 1-figure supplement 1). 136 6 137 To ensure that mutation of Asp2025.42 did not impair protein folding or G protein coupling, we 138 asked whether the mutant receptor could be activated by other ligands. We tested 50 chemicals 139 for the ability to activate TAAR13cD202A, and identified the monoamine 3-methoxytyramine as an 140 agonist (Figure 1D). 3-methoxytyramine failed to activate wild type TAAR13c at any 141 concentration tested.
Recommended publications
  • Impact of Using Organic Yeast in the Fermentation Process of Wine
    processes Article Impact of Using Organic Yeast in the Fermentation Process of Wine Balázs Nagy 1, Zsuzsanna Varga 2,Réka Matolcsi 1, Nikolett Kellner 1 , Áron Szövényi 1 and Diána Nyitrainé Sárdy 1,* 1 Faculty of Horticultural Science Department of Oenology, Szent István University, 1118 Budapest, Hungary; [email protected] (B.N.); [email protected] (R.M.); [email protected] (N.K.); [email protected] (Á.S.) 2 Faculty of Horticultural Science Department of Viticulture, Szent István University, 1118 Budapest, Hungary; [email protected] * Correspondence: [email protected] Abstract: The aim of this study was to find out what kind of “Bianca” wine could be produced when using organic yeast, what are the dynamics of the resulting alcoholic fermentation, and whether this method is suitable for industrial production as well. Due to the stricter rules and regulations, as well as the limited amount and selection of the permitted chemicals, resistant, also known as interspecific or innovative grape varieties, can be the ideal basic materials of alternative cultivation technologies. Well-designed analytical and organoleptic results have to provide the scientific background of resistant varieties, as these cultivars and their environmentally friendly cultivation techniques could be the raw materials of the future. The role of the yeast in wine production is crucial. We fermented wines from the “Bianca” juice samples three times where model chemical solutions were applied. In our research, we aimed to find out how organic yeast influenced the biogenic amine formation of three important compounds: histamine, tyramine, and serotonin. The main results of this study showed that all the problematic values (e.g., histamine) were under the critical limit (1 g/L), although the organic samples resulted in a significantly higher level than the control wines.
    [Show full text]
  • Transport of Dangerous Goods
    ST/SG/AC.10/1/Rev.16 (Vol.I) Recommendations on the TRANSPORT OF DANGEROUS GOODS Model Regulations Volume I Sixteenth revised edition UNITED NATIONS New York and Geneva, 2009 NOTE The designations employed and the presentation of the material in this publication do not imply the expression of any opinion whatsoever on the part of the Secretariat of the United Nations concerning the legal status of any country, territory, city or area, or of its authorities, or concerning the delimitation of its frontiers or boundaries. ST/SG/AC.10/1/Rev.16 (Vol.I) Copyright © United Nations, 2009 All rights reserved. No part of this publication may, for sales purposes, be reproduced, stored in a retrieval system or transmitted in any form or by any means, electronic, electrostatic, magnetic tape, mechanical, photocopying or otherwise, without prior permission in writing from the United Nations. UNITED NATIONS Sales No. E.09.VIII.2 ISBN 978-92-1-139136-7 (complete set of two volumes) ISSN 1014-5753 Volumes I and II not to be sold separately FOREWORD The Recommendations on the Transport of Dangerous Goods are addressed to governments and to the international organizations concerned with safety in the transport of dangerous goods. The first version, prepared by the United Nations Economic and Social Council's Committee of Experts on the Transport of Dangerous Goods, was published in 1956 (ST/ECA/43-E/CN.2/170). In response to developments in technology and the changing needs of users, they have been regularly amended and updated at succeeding sessions of the Committee of Experts pursuant to Resolution 645 G (XXIII) of 26 April 1957 of the Economic and Social Council and subsequent resolutions.
    [Show full text]
  • The Histamine H4 Receptor: a Novel Target for Safe Anti-Inflammatory
    GASTRO ISSN 2377-8369 Open Journal http://dx.doi.org/10.17140/GOJ-1-103 Review The Histamine H4 Receptor: A Novel Target *Corresponding author Maristella Adami, PhD for Safe Anti-inflammatory Drugs? Department of Neuroscience University of Parma Via Volturno 39 43125 Parma Italy * 1 Tel. +39 0521 903943 Maristella Adami and Gabriella Coruzzi Fax: +39 0521 903852 E-mail: [email protected] Department of Neuroscience, University of Parma, Via Volturno 39, 43125 Parma, Italy Volume 1 : Issue 1 1retired Article Ref. #: 1000GOJ1103 Article History Received: May 30th, 2014 ABSTRACT Accepted: June 12th, 2014 th Published: July 16 , 2014 The functional role of histamine H4 receptors (H4Rs) in the Gastrointestinal (GI) tract is reviewed, with particular reference to their involvement in the regulation of gastric mucosal defense and inflammation. 4H Rs have been detected in different cell types of the gut, including Citation immune cells, paracrine cells, endocrine cells and neurons, from different animal species and Adami M, Coruzzi G. The Histamine H4 Receptor: a novel target for safe anti- humans; moreover, H4R expression was reported to be altered in some pathological conditions, inflammatory drugs?. Gastro Open J. such as colitis and cancer. Functional studies have demonstrated protective effects of H4R an- 2014; 1(1): 7-12. doi: 10.17140/GOJ- tagonists in several experimental models of gastric mucosal damage and intestinal inflamma- 1-103 tion, suggesting a potential therapeutic role of drugs targeting this new receptor subtype in GI disorders, such as allergic enteropathy, Inflammatory Bowel Disease (IBD), Irritable Bowel Syndrome (IBS) and cancer. KEYWORDS: Histamine H4 receptor; Stomach; Intestine.
    [Show full text]
  • Emerging Evidence for a Central Epinephrine-Innervated A1- Adrenergic System That Regulates Behavioral Activation and Is Impaired in Depression
    Neuropsychopharmacology (2003) 28, 1387–1399 & 2003 Nature Publishing Group All rights reserved 0893-133X/03 $25.00 www.neuropsychopharmacology.org Perspective Emerging Evidence for a Central Epinephrine-Innervated a1- Adrenergic System that Regulates Behavioral Activation and is Impaired in Depression ,1 1 1 1 1 Eric A Stone* , Yan Lin , Helen Rosengarten , H Kenneth Kramer and David Quartermain 1Departments of Psychiatry and Neurology, New York University School of Medicine, New York, NY, USA Currently, most basic and clinical research on depression is focused on either central serotonergic, noradrenergic, or dopaminergic neurotransmission as affected by various etiological and predisposing factors. Recent evidence suggests that there is another system that consists of a subset of brain a1B-adrenoceptors innervated primarily by brain epinephrine (EPI) that potentially modulates the above three monoamine systems in parallel and plays a critical role in depression. The present review covers the evidence for this system and includes findings that brain a -adrenoceptors are instrumental in behavioral activation, are located near the major monoamine cell groups 1 or target areas, receive EPI as their neurotransmitter, are impaired or inhibited in depressed patients or after stress in animal models, and a are restored by a number of antidepressants. This ‘EPI- 1 system’ may therefore represent a new target system for this disorder. Neuropsychopharmacology (2003) 28, 1387–1399, advance online publication, 18 June 2003; doi:10.1038/sj.npp.1300222 Keywords: a1-adrenoceptors; epinephrine; motor activity; depression; inactivity INTRODUCTION monoaminergic systems. This new system appears to be impaired during stress and depression and thus may Depressive illness is currently believed to result from represent a new target for this disorder.
    [Show full text]
  • Ligand-Gated Chloride Channels Are Receptors for Biogenic Amines in C
    Ligand-Gated Chloride Channels Are Receptors for Biogenic Amines in C. elegans The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation Ringstad, N., N. Abe, and H. R. Horvitz. “Ligand-Gated Chloride Channels Are Receptors for Biogenic Amines in C. elegans.” Science 325, no. 5936 (July 2, 2009): 96-100. As Published http://dx.doi.org/10.1126/science.1169243 Publisher American Association for the Advancement of Science (AAAS) Version Author's final manuscript Citable link http://hdl.handle.net/1721.1/84506 Terms of Use Creative Commons Attribution-Noncommercial-Share Alike 3.0 Detailed Terms http://creativecommons.org/licenses/by-nc-sa/3.0/ NIH Public Access Author Manuscript Science. Author manuscript; available in PMC 2010 October 25. NIH-PA Author ManuscriptPublished NIH-PA Author Manuscript in final edited NIH-PA Author Manuscript form as: Science. 2009 July 3; 325(5936): 96±100. doi:10.1126/science.1169243. Ligand-gated chloride channels are receptors for biogenic amines in C. elegans Niels Ringstad1,2, Namiko Abe1,2,3, and H. Robert Horvitz1 1HHMI, Department of Biology and McGovern Institute for Brain Research, MIT, Cambridge MA 02139 Abstract Biogenic amines such as serotonin and dopamine are intercellular signaling molecules that function widely as neurotransmitters and neuromodulators. We have identified in the nematode Caenorhabditis elegans three ligand-gated chloride channels that are receptors for biogenic amines: LGC-53 is a high-affinity dopamine receptor, LGC-55 is a high-affinity tyramine receptor, and LGC-40 is a low-affinity serotonin receptor that is also gated by choline and acetylcholine.
    [Show full text]
  • 4695389.Pdf (3.200Mb)
    Non-classical amine recognition evolved in a large clade of olfactory receptors The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Li, Qian, Yaw Tachie-Baffour, Zhikai Liu, Maude W Baldwin, Andrew C Kruse, and Stephen D Liberles. 2015. “Non-classical amine recognition evolved in a large clade of olfactory receptors.” eLife 4 (1): e10441. doi:10.7554/eLife.10441. http://dx.doi.org/10.7554/ eLife.10441. Published Version doi:10.7554/eLife.10441 Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:23993622 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA RESEARCH ARTICLE Non-classical amine recognition evolved in a large clade of olfactory receptors Qian Li1, Yaw Tachie-Baffour1, Zhikai Liu1, Maude W Baldwin2, Andrew C Kruse3, Stephen D Liberles1* 1Department of Cell Biology, Harvard Medical School, Boston, United States; 2Department of Organismic and Evolutionary Biology, Museum of Comparative Zoology, Harvard University, Cambridge, United States; 3Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States Abstract Biogenic amines are important signaling molecules, and the structural basis for their recognition by G Protein-Coupled Receptors (GPCRs) is well understood. Amines are also potent odors, with some activating olfactory trace amine-associated receptors (TAARs). Here, we report that teleost TAARs evolved a new way to recognize amines in a non-classical orientation.
    [Show full text]
  • Decarboxylation What Is Decarboxylation?
    Decarboxylation What is decarboxylation? Decarboxylation is the removal of a carboxyl group when a compound is exposed to light or heat. A carboxyl group is a grouping of carbon, oxygen, and hydrogen atoms in the form of COOH. When a compound is decarboxylated, this group is released in the form of carbon dioxide. Decarboxylation of cannabinoids When considering the decarboxylation of cannabinoids, it is important to note that this reaction is what causes the neutral acidic cannabinoids to turn into the active cannabinoids found in most products on the market. Both the neutral and acidic cannabinoids are naturally found in the cannabis plant. The active cannabinoids that have psychoactive properties are what occur after decarboxylation. Below is the mechanism for the conversion of THCA to THC. https://www.leafscience.com/2017/04/13/what-is-thca/ Heating THCA converts it to THC by kicking off the carboxyl group seen on the right side of the THCA molecule. Once the carboxyl group is released from the THCA it forms carbon dioxide and the THC compound. The THC is now ready for use in different products and will provide the psychoactive effects that popularized this compound. This simple reaction is all it takes to convert the acidic cannabinoids to their active counterparts. Cannabinolic acid (CBDA) and CBD are both another example of this process. Decarboxylation process Light can start the decarboxylation process after a period of time, but heat is the more common element used. There are different methods to decarboxylate cannabis depending on user preference. In the industry, dabs, vapes and joints decarboxylate instantly when heat is applied to the product.
    [Show full text]
  • Neurotransmitters-Drugs Andbrain Function.Pdf
    Neurotransmitters, Drugs and Brain Function. Edited by Roy Webster Copyright & 2001 John Wiley & Sons Ltd ISBN: Hardback 0-471-97819-1 Paperback 0-471-98586-4 Electronic 0-470-84657-7 Neurotransmitters, Drugs and Brain Function Neurotransmitters, Drugs and Brain Function. Edited by Roy Webster Copyright & 2001 John Wiley & Sons Ltd ISBN: Hardback 0-471-97819-1 Paperback 0-471-98586-4 Electronic 0-470-84657-7 Neurotransmitters, Drugs and Brain Function Edited by R. A. Webster Department of Pharmacology, University College London, UK JOHN WILEY & SONS, LTD Chichester Á New York Á Weinheim Á Brisbane Á Singapore Á Toronto Neurotransmitters, Drugs and Brain Function. Edited by Roy Webster Copyright & 2001 John Wiley & Sons Ltd ISBN: Hardback 0-471-97819-1 Paperback 0-471-98586-4 Electronic 0-470-84657-7 Copyright # 2001 by John Wiley & Sons Ltd. Bans Lane, Chichester, West Sussex PO19 1UD, UK National 01243 779777 International ++44) 1243 779777 e-mail +for orders and customer service enquiries): [email protected] Visit our Home Page on: http://www.wiley.co.uk or http://www.wiley.com All Rights Reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording, scanning or otherwise, except under the terms of the Copyright, Designs and Patents Act 1988 or under the terms of a licence issued by the Copyright Licensing Agency Ltd, 90 Tottenham Court Road, London W1P0LP,UK, without the permission in writing of the publisher. Other Wiley Editorial Oces John Wiley & Sons, Inc., 605 Third Avenue, New York, NY 10158-0012, USA WILEY-VCH Verlag GmbH, Pappelallee 3, D-69469 Weinheim, Germany John Wiley & Sons Australia, Ltd.
    [Show full text]
  • Optimization of the Decarboxylation Reaction in Cannabis
    APPLICATION NOTE FT-IR Spectroscopy Authors: Dr. Markus Roggen1 Antonio M. Marelli1 Doug Townsend2 Ariel Bohman2 1Outco SanDiego, CA 2PerkinElmer, Inc. Shelton, CT. Optimization of the Decarboxylation Reaction Introduction The production of cannabis in Cannabis Extract extracts and oils for medicinal and recreational products has increased significantly in North America. This growth has been driven by both market demand in newly legalized states and patient demand for a greater diversity in cannabis products.1,2,3 Most cannabis extraction processes, independent of solvent or instrument choice, undergo a decarboxylation step whereby the carboxylic acid functional group is removed from the cannabinoids. The decarboxylation reaction converts the naturally occurring acid forms of the cannabinoids, e.g. tetrahydrocannabinolic acid (THCA) and cannabidiolic acid (CBDA), to their more potent neutral forms, e.g. tetrahydrocannabinol (THC) and cannabidiol (CBD). Because the carboxylic acid group is thermally labile, the industry typically applies a heat source, and at times a catalyst, to decarboxylate the cannabinoids. The heat-promoted decarboxylation reaction has been Decarboxylation Reaction discussed at length within the industry, but an extensive Decarboxylation was achieved by heating the cannabis extract in 4, 5, 6 literature search reveals very few papers on the process. a oil bath with a programmable hot plate. An overhead stirrer The data available represents a large spectrum of reaction was utilized to promote even heat distribution throughout the conditions, including a range in reaction temperature, time experiment. Oil bath and extract temperatures were recorded and instrumental setup. As such, there is a lack of universal every five minutes throughout the 80-minute heating process.
    [Show full text]
  • The Role of Inflammatory Pathways in Neuroblastoma Tumorigenesis — Igor Snapkov a Dissertation for the Degree of Philosophiae Doctor – August 2016 Contents
    Faculty of Health Sciences Department of Medical Biology Molecular Inflammation Research Group The role of inflammatory pathways in neuroblastoma tumorigenesis — Igor Snapkov A dissertation for the degree of Philosophiae Doctor – August 2016 Contents 1. List of publications .................................................................................................................. 2 2. List of abbreviations ................................................................................................................ 3 3. Introduction ............................................................................................................................. 5 3.1 Cancer and inflammation .................................................................................................. 5 3.2.1 Pattern recognition receptors and danger signals ............................................................ 7 3.2.2 Formyl peptide receptor 1 (FPR1) ................................................................................. 9 3.3.1 Chemokines................................................................................................................. 12 3.3.2 Chemerin ..................................................................................................................... 13 3.4 Neuroblastoma ................................................................................................................ 14 3.5 Hepatic clearance of danger signals ................................................................................
    [Show full text]
  • Biogenic Amines Formation and Their Importance in Fermented Foods
    BIO Web of Conferences 17, 00232 (2020) https://doi.org/10.1051/bioconf/20201700232 FIES 2019 Biogenic amines formation and their importance in fermented foods Kamil Ekici1, ⃰ and Abdullah Khalid Omer2 1University of Van Yȕzȕncȕ Yıl, Veterinary College, Department of Food Hygiene and Technology, Van, Turkey 2Sulaimani Veterinary Directorate, Veterinary Quarantine, Bashmakh International Border, Sulaimani, Iraq Abstract. Biogenic amines (BAs) are low molecular weight organic bases with an aliphatic, aromatic, or heterocyclic structure which have been found in many foods. biogenic amines have been related with several outbreaks of food-borne intoxication and are very important in public health concern because of their potential toxic effects. The accumulation of biogenic amines in foods is mainly due to the presence of bacteria able to decarboxylate certain amino acids. Biogenic amines are formed when the alpha carboxvl group breaks away from free amino acid precursors. They are colled after the amino acid they originated from. The main biogenic amines producers in foods are Gram positive bacteria and cheese is among the most commonly implicated foods associated with biogenic amines poisoning. The consumption of foods containing high concentrations of biogenic amines has been associated with health hazards and they are used as a quality indicator that shows the degree of spoilage, use of non-hygienic raw material and poor manufacturing practice. Biogenic amines may also be considered as carcinogens because they are able to react with nitrites to form potentially carcinogenic nitrosamines. Generally, biogenic amines in foods can be controlled by strict use of good hygiene in both raw material and manufacturing environments with corresponding inhibition of spoiling microorganisms.
    [Show full text]
  • Histamine Receptors
    Tocris Scientific Review Series Tocri-lu-2945 Histamine Receptors Iwan de Esch and Rob Leurs Introduction Leiden/Amsterdam Center for Drug Research (LACDR), Division Histamine is one of the aminergic neurotransmitters and plays of Medicinal Chemistry, Faculty of Sciences, Vrije Universiteit an important role in the regulation of several (patho)physiological Amsterdam, De Boelelaan 1083, 1081 HV, Amsterdam, The processes. In the mammalian brain histamine is synthesised in Netherlands restricted populations of neurons that are located in the tuberomammillary nucleus of the posterior hypothalamus.1 Dr. Iwan de Esch is an assistant professor and Prof. Rob Leurs is These neurons project diffusely to most cerebral areas and have full professor and head of the Division of Medicinal Chemistry of been implicated in several brain functions (e.g. sleep/ the Leiden/Amsterdam Center of Drug Research (LACDR), VU wakefulness, hormonal secretion, cardiovascular control, University Amsterdam, The Netherlands. Since the seventies, thermoregulation, food intake, and memory formation).2 In histamine receptor research has been one of the traditional peripheral tissues, histamine is stored in mast cells, eosinophils, themes of the division. Molecular understanding of ligand- basophils, enterochromaffin cells and probably also in some receptor interaction is obtained by combining pharmacology specific neurons. Mast cell histamine plays an important role in (signal transduction, proliferation), molecular biology, receptor the pathogenesis of various allergic conditions. After mast cell modelling and the synthesis and identification of new ligands. degranulation, release of histamine leads to various well-known symptoms of allergic conditions in the skin and the airway system. In 1937, Bovet and Staub discovered compounds that antagonise the effect of histamine on these allergic reactions.3 Ever since, there has been intense research devoted towards finding novel ligands with (anti-) histaminergic activity.
    [Show full text]