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PMID: 1847522 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

gamma-Aminobutyric acid type A receptor point mutation increases the affinity of compounds for the benzodiazepine site.

Pritchett DB, Seeburg PH

Abstract

Recombinantly expressed gamma-aminobutyric acid type A (GABAA) receptors consisting of alpha 1, beta 2, and gamma 2 subunits contain a binding site for benzodiazepines that differs in its properties from that of alpha 3 beta 2 gamma 2 receptors. Amino acid substitutions between the GABAA receptor alpha subunits were analyzed for their effect on the binding of compounds to the benzodiazepine site. By converting ever smaller regions of the alpha 3 subunit sequence to that of the alpha 1 subunit, we show that a single substitution (glycine for glutamic acid) increases the affinity for several compounds approximately 10-fold without changing the affinity for nonselective compounds. Hence, the identified amino acids may interact directly with the ligand and define part of the benzodiazepine binding sites in these receptors.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites Binding, Competitive Cell Line Chimera Flumazenil/metabolism Humans Kinetics Macromolecular Substances Molecular Sequence Data Mutagenesis, Site-Directed Oligonucleotide Probes Rats Receptors, GABA-A/genetics,metabolism Transfection
Chemicals
Macromolecular Substances Oligonucleotide Probes Receptors, GABA-A Flumazenil
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Pritchett D B
Department of Pediatrics, University of Pennsylvania, Philadelphia 19104.
Seeburg P H
References (27)
27 references, click to expand
  1. Structural and functional characterization of the gamma 1 subunit of GABAA/benzodiazepine receptors.
    EMBO J. 1990 Oct;9(10):3261-7 PMID: 2170110
  2. Oligonucleotide-directed mutagenesis using M13-derived vectors: an efficient and general procedure for the production of point mutations in any fragment of DNA.
    Nucleic Acids Res. 1982 Oct 25;10(20):6487-500 PMID: 6757864
  3. Acetylcholine receptor binding site contains a disulfide cross-link between adjacent half-cystinyl residues.
    J Biol Chem. 1986 Jun 25;261(18):8085-8 PMID: 3722144
  4. High-efficiency transformation of mammalian cells by plasmid DNA.
    Mol Cell Biol. 1987 Aug;7(8):2745-52 PMID: 3670292
  5. Transient expression shows ligand gating and allosteric potentiation of GABAA receptor subunits.
    Science. 1988 Dec 2;242(4883):1306-8 PMID: 2848320
  6. A synthetic non-benzodiazepine ligand for benzodiazepine receptors: a probe for investigating neuronal substrates of anxiety.
    Pharmacol Biochem Behav. 1979 Jul;11(1):99-106 PMID: 40260
  7. Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction.
    Biochem Pharmacol. 1973 Dec 1;22(23):3099-108 PMID: 4202581
  8. Evidence for involvement of GABA in the action of benzodiazepines: studies on rat cerebellum.
    Adv Biochem Psychopharmacol. 1975;(14):113-30 PMID: 242198
  9. Structural and functional basis for GABAA receptor heterogeneity.
    Nature. 1988 Sep 1;335(6185):76-9 PMID: 2842688
  10. Transmembrane topology and subcellular distribution of the benzodiazepine receptor.
    J Neurosci. 1986 Oct;6(10):2857-63 PMID: 3020185
  11. Pharmacological profile of the imidazopyridine zolpidem at benzodiazepine receptors and electrocorticogram in rats.
    Naunyn Schmiedebergs Arch Pharmacol. 1985 Sep;330(3):248-51 PMID: 2997631
  12. Differences in the negative allosteric modulation of gamma-aminobutyric acid receptors elicited by 4'-chlorodiazepam and by a beta-carboline-3-carboxylate ester: a study with natural and reconstituted receptors.
    Proc Natl Acad Sci U S A. 1989 Sep;86(18):7275-9 PMID: 2476816
  13. An analog of lophotoxin reacts covalently with Tyr190 in the alpha-subunit of the nicotinic acetylcholine receptor.
    J Biol Chem. 1989 Jul 25;264(21):12666-72 PMID: 2568359
  14. A novel alpha subunit in rat brain GABAA receptors.
    Neuron. 1989 Dec;3(6):745-53 PMID: 2561977
  15. Two novel GABAA receptor subunits exist in distinct neuronal subpopulations.
    Neuron. 1989 Sep;3(3):327-37 PMID: 2561970
  16. Sequence and expression of a novel GABAA receptor alpha subunit.
    FEBS Lett. 1989 Nov 20;258(1):119-22 PMID: 2556293
  17. GABAA receptor beta subunit heterogeneity: functional expression of cloned cDNAs.
    EMBO J. 1989 Jun;8(6):1665-70 PMID: 2548852
  18. Type I and type II GABAA-benzodiazepine receptors produced in transfected cells.
    Science. 1989 Sep 22;245(4924):1389-92 PMID: 2551039
  19. Importance of a novel GABAA receptor subunit for benzodiazepine pharmacology.
    Nature. 1989 Apr 13;338(6216):582-5 PMID: 2538761
  20. Transmembrane topology of the gamma-aminobutyric acidA/benzodiazepine receptor: subcellular distribution and allosteric coupling determined in situ.
    Mol Pharmacol. 1989 Jan;35(1):75-84 PMID: 2536471
  21. An LFA-3 cDNA encodes a phospholipid-linked membrane protein homologous to its receptor CD2.
    Nature. 1987 Oct 29-Nov 4;329(6142):840-2 PMID: 3313052
  22. Cerebellar GABAA receptor selective for a behavioural alcohol antagonist.
    Nature. 1990 Aug 16;346(6285):648-51 PMID: 2166916
  23. Functional properties of recombinant rat GABAA receptors depend upon subunit composition.
    Neuron. 1990 Jun;4(6):919-28 PMID: 1694446
  24. Gamma-aminobutyric acidA receptor alpha 5-subunit creates novel type II benzodiazepine receptor pharmacology.
    J Neurochem. 1990 May;54(5):1802-4 PMID: 2157817
  25. Several new benzodiazepines selectively interact with a benzodiazepine receptor subtype.
    Neurosci Lett. 1983 Jul 15;38(1):73-8 PMID: 6136944
  26. Ethyl beta-carboline-3-carboxylate shows differential benzodiazepine receptor interaction.
    Nature. 1980 Aug 7;286(5773):606-7 PMID: 6250077
  27. Chimeric alpha 2-,beta 2-adrenergic receptors: delineation of domains involved in effector coupling and ligand binding specificity.
    Science. 1988 Jun 3;240(4857):1310-6 PMID: 2836950
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1991-02-15
Pages
1421-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC51030
Subset
IM
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