Cysteinyl leukotriene receptors

doi:10.1016/S0006-2952(02)01357-6

Biochemical Pharmacology

Volume 64, Issue 11, 1 December 2002, Pages 1549-1557

https://doi.org/10.1016/S0006-2952(02)01357-6 Get rights and content

Abstract

Cysteinyl leukotrienes (CysLTs) are important inflammatory mediators in asthma and allergic disorders. Two types of CysLT receptors, CysLT₁ and CysLT₂, which were originally defined pharmacologically based on their sensitivity to CysLT₁ specific antagonists, are responsible for most of the known CysLT biological actions. The regulation of CysLT receptor expression and signaling in disease processes is largely unclear. Recent molecular cloning of both receptor subtypes from several different species will greatly facilitate future research in understanding CysLT signal transduction mechanisms. Expression of the relatively better-studied CysLT₁ is verified in lung tissues and peripheral blood cells. Elucidating how this receptor mediates airway inflammation will deepen our understanding of asthma etiology. On the other hand, detection of CysLT₂ in the heart, brain, and adrenal glands will inject new excitement into the search for novel CysLT functions. This review summarizes receptor cloning, ligand binding, expression, signaling, and functions in an effort to bridge early pharmacological studies to future studies at the molecular level.

Introduction

The study of the CysLTs LTC₄, LTD₄, and LTE₄, known historically as slow-reacting substance of anaphylaxis (SRS-A), has been ongoing for more than twenty years [1]. CysLTs are important inflammatory mediators that play a major role in the pathophysiology of inflammatory diseases such as asthma and allergic rhinitis.

CysLTs exert their effects through cell surface receptors. Great efforts have been put into the characterization of the receptors ever since the discovery of these substances. In fact, FPL-55712 was identified as a specific antagonist even before the chemical components of SRS-A were elucidated [2]. Significant confusion in receptor classification existed prior to the standardization of the nomenclature in 1995 by the International Union of Pharmacologists (IUPHAR) [3]. CysLT receptors were classified into two major subtypes: CysLT₁, which could be blocked by a family of specific antagonists, and CysLT₂, which was basically insensitive to CysLT₁ antagonists. This classification, however, did not rule out the possibility of subdivision. Recently, the molecular cloning of both CysLT₁[4], [5], [6], [7] and CysLT₂[8], [9], [10], [11] has opened new avenues for CysLT exploration.

Section snippets

CysLT receptor cDNA and gene cloning

Numerous quests at conventional receptor purification or expression cloning proved futile for cloning CysLT receptors. Their ultimate cloning resulted from blooming sequence databases, computational mining, and high throughput screening methodologies. hCysLT₁ cDNA encodes a protein of 337 amino acids (aa). mCysLT₁ is encoded by two transcripts: a short splice variant encoding a protein of 339 aa, which aligns well with hCysLT₁; and a long variant encoding a protein extending 13 aa at the N

Protein structure and ligand binding of CysLT receptors

CysLT receptors have long been recognized as GPCRs prior to their cloning [13], based on the fact that binding of the ligands to the receptors is enhanced by divalent cations but inhibited by sodium ions and non-hydrolyzable GTP analogs.

Hydrophobicity analysis of the deduced primary structures reveals that both CysLT₁ and CysLT₂ have seven hydrophobic transmembrane (TM) domains linked by six hydrophilic loops, typical of GPCRs. There is no binding model currently available for leukotrienes.

Signaling through CysLT receptors

In each of the reported cloning studies, intracellular calcium mobilization was used to measure receptor function in vitro. In the cells transfected with CysLT₁ cDNA, CysLT could increase Ca²⁺ concentration in a rank order potency of LTD₄>LTC₄≥LTE₄[4], [5], [6], [7]; or LTC₄=LTD₄>LTE₄ in cells transfected with CysLT₂ cDNA [8], [9], [10], [11]. This is consistent with pharmacological studies described previously [3]. Species variations have been described in different studies [6], [7], [19], [20]

Expression and regulation of CysLT receptors

CysLT receptor tissue distribution has classically been linked to binding studies or functional characterization, using tissue preparations, by observing CysLT effects and the reversing actions of CysLT₁ specific antagonists or the CysLT_1/2 dual antagonist BAYu9773. Considering CysLTs as inflammatory mediators and their significant contributions to human asthma, expression patterns were characterized in lung and inflammatory cells [32]. CysLT₁ and CysLT₂ expression patterns vary considerably in

Functions of CysLT receptors

The role of CysLTs in inflammation has been studied intensively, and emphasis has been placed on human asthma. A large market for antileukotriene drugs to treat human asthma symptoms has been successfully established since 1995. CysLT and their receptor function studies are not confined to asthma etiology. In fact, the concept that CysLTs are mainly inflammatory mediators may be modified as evidence of their potential involvement in cardiovascular and neurological modulation is accumulating.

Prospects

Since their discovery, CysLTs have been regarded mainly as important mediators of inflammation, and research accordingly has been tied to the elucidation of their effects in inflammatory processes. The molecular cloning of the two CysLT receptor subtypes has not only confirmed many previous pharmacological observations but has also broadened the horizon to unexpected physiological effects of CysLTs beyond inflammation and removed a hurdle to elucidate CysLT signal transduction mechanisms. How

Acknowledgements

This work was supported by NIH Grant HL58464.

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These cytokines, in turn, stimulate the secretion of more cytokines, increase adhesion molecule expression by eosinophils, and promote eosinophil survival through CysLT1.167,168 CysLT1 is believed to be responsible for the majority of CysLT-mediated effects in asthma because such effects can be reversed by treatment with CysLT1 antagonists.169 A role of CysLTs/CysLT1 in the clinical manifestations of asthma is supported by the finding that LTRAs (montelukast, zafirlukast, and pranlukast) are an effective treatment for asthma, in addition to the 5-LO inhibitor, zileuton.149
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CommentaryCysteinyl leukotriene receptors

Abstract

Introduction

Section snippets

CysLT receptor cDNA and gene cloning

Protein structure and ligand binding of CysLT receptors

Signaling through CysLT receptors

Expression and regulation of CysLT receptors

Functions of CysLT receptors

Prospects

Acknowledgements

Biochem. Pharmacol.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Trends Genet.

Eur. J. Pharmacol.

J. Biol. Chem.

Eur. J. Pharmacol.

Eur. J. Pharmacol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Trends Pharmacol. Sci.

Pulm. Pharmacol. Ther.

Prostaglandins Leukot. Essent. Fatty Acids

Biochem. Biophys. Res. Commun.

Am. J. Pathol.

Chest

J. Allergy Clin. Immunol.

J. Allergy Clin. Immunol.

Lancet

Pulm. Pharmacol.

Cell

J. Am. Coll. Cardiol.

Eur. J. Pharmacol.

FEBS Lett.

Eur. J. Pharmacol.

Brain Res.

Leukotriene C: a slow-reacting substance from murine mastocytoma cells

Proc. Natl. Acad. Sci. U.S.A.

Selective inhibitor of slow reacting substance of anaphylaxis

Nat. New Biol.

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Adv. Prostaglandin Thromboxane Leukot. Res.

Characterization of the human cysteinyl leukotriene CysLT1 receptor

Nature

Identification, molecular cloning, expression, and characterization of a cysteinyl leukotriene receptor

Mol. Pharmacol.

Identification in mice of two isoforms of the cysteinyl leukotriene 1 receptor that result from alternative splicing

Proc. Natl. Acad. Sci. U.S.A.

Molecular cloning and characterization of a second human cysteinyl leukotriene receptor: discovery of a subtype selective agonist

Mol. Pharmacol.

Point mutation in the seventh hydrophobic domain of the human thromboxane A2 receptor allows discrimination between agonist and antagonist binding sites

Mol. Pharmacol.

Cysteinyl leukotriene receptor 1 is also a pyrimidinergic receptor and is expressed by human mast cells

Proc. Natl. Acad. Sci. U.S.A.

Human P2Y1 receptor: molecular modeling and site-directed mutagenesis as tools to identify agonist and antagonist recognition sites

J. Med. Chem.

Role of the extracellular loops of G protein-coupled receptors in ligand recognition: a molecular modeling study of the human P2Y1 receptor

Biochemistry

Do recombinant receptor assays provide affinity and potency estimates?

Ann. N.Y. Acad. Sci.

Leukotriene D4 receptor-mediated phosphoinositol hydrolysis and calcium mobilization in rat basophilic leukemic cells

J. Pharmacol. Exp. Ther.

Leukotriene D4 receptor-mediated hydrolysis of phosphoinositide and mobilization of calcium in sheep tracheal smooth muscle cells

J. Pharmacol. Exp. Ther.

Transient activation of topoisomerase I in leukotriene D4 signal transduction in human cells

Biochem. J.

Structural and functional aspects of G protein-coupled receptor oligomerization

Biochem. Cell Biol.

Dimerization of G-protein-coupled receptors

J. Med. Chem.

Expression of the cysteinyl leukotriene 1 receptor in normal human lung and peripheral blood leukocytes

Am. J. Respir. Crit. Care Med.

Binding to cysteinyl-leukotriene receptors

Am. J. Respir. Crit. Care Med.

Pharmacological characterization of leukotriene receptors

Am. J. Respir. Crit. Care Med.

Levels of cysteinyl leukotriene receptor mRNA in human peripheral leucocytes: significantly higher expression of cysteinyl leukotriene receptor 2 mRNA in eosinophils

Commentary
Cysteinyl leukotriene receptors

Characterization of the human cysteinyl leukotriene CysLT₁ receptor

Point mutation in the seventh hydrophobic domain of the human thromboxane A₂ receptor allows discrimination between agonist and antagonist binding sites

Leukotriene D₄ receptor-mediated phosphoinositol hydrolysis and calcium mobilization in rat basophilic leukemic cells

Leukotriene D₄ receptor-mediated hydrolysis of phosphoinositide and mobilization of calcium in sheep tracheal smooth muscle cells

Transient activation of topoisomerase I in leukotriene D₄ signal transduction in human cells