Prostaglandins and Leukotrienes

W L Smith, University of Michigan Medical School, Ann Arbor, MI, USA

R C Murphy, University of Colorado Medical School, Denver, CO, USA
ã 2013 Elsevier Inc. All rights reserved.

This article is reproduced from the previous edition, volume 3, pp. 452–456, ã 2004, Elsevier Inc.

Glossary Leukotriene Those oxygenated derivatives of 20 carbon

5-Lipoxygenase The oxygenase that converts arachidonic essential fatty acids formed through the action of
acid and related 20 carbon essential fatty acids to leukotrienes. 5-lipoxygenase and containing a conjugated triene system of
Aspirin A nonsteroidal anti-inflammatory drug that inhibits double bonds.
cyclooxygenase by acetylating an active site serine residue Montelukast A drug used in the treatment of asthma that
and blocking productive binding of arachidonic acid. inhibits the binding of peptidoleukotrienes to the CysLT1
Cyclooxygenase Common name of the oxygenase enzyme receptor.
that converts arachidonic acid and related 20 carbon Prostanoid Those oxygenated derivatives of 20 carbon
essential fatty acids to prostanoids. essential fatty acids formed through the action of
FLAP 5-Lipoxygenase activating protein, an accessory protein prostaglandin endoperoxide H synthase.
that is required for 5-lipoxygenase functioning in intact cells.

Prostanoid Structures (e.g., TXA synthase and PGE synthase). Recently, 2-AG has
been found to be converted to 2-PGH2-glycerol by PGHS-2.
Figure 1 shows the biosynthetic relationships among the pros- This latter intermediate can be converted to the 2-prostanyl-
tanoids formed from AA. The letters following the abbreviation glycerol derivatives but not 2-thromboxane-glycerol. 2-AG
PGs indicate the nature and location of the oxygen-containing itself is probably formed from phosphatidylcholine by phos-
substituents present in the cyclopentane ring; the subscript pholipase C and monoacylglycerol lipase.
indicates the number of carbon–carbon double bonds in the
side chains. Greek subscripts denote the orientation of ring
PGHS Catalysis and Inhibition
hydroxyl groups (e.g., PGF2a). Prostanoids known as isopros-
tanes are formed from AA by nonenzymatic peroxidation and
Figure 2 is a model of the COX and peroxidase active sites of
consequently contain many stereoisomers. Interestingly, the
ovine PGHS-1. To initiate oxygenation of AA at the COX site, the
amounts of isoprostane metabolites in urine are greater than
heme group at the peroxidase site must first be oxidized by a
prostanoids formed enzymatically.
hydroperoxide. This causes formation of an oxidized heme
intermediate that is reduced by an electron from Tyr385, thereby
Prostanoid Biosynthesis generating a Tyr385 radical. This radical abstracts a hydrogen
from AA so that it can react with oxygen (Figure 1). The guani-
Eicosanoids are not stored in cells but are formed on demand in dino group of Arg120 binds to the carboxylate group of AA.
response to extracellular hormonal stimuli (e.g., bradykinin, PGHS-1 is a constitutive enzyme purified in 1975 and its
angiotensin, and thrombin) that increase cell Ca2þ concentra- cDNA cloning has been reported in 1988. PGHS-2 is an induc-
tions. Prostanoid formation requires three enzymatic steps: ible enzyme discovered in 1991 as an immediate early gene
(1) mobilization of AA (or 2-arachidonylglycerol (2-AG)) from product in phorbol ester-activated murine 3T3 cells and in
membrane phospholipids; (2) conversion of arachidonate (or v-src-transformed chicken fibroblasts. PGHS-2 contains a
2-AG) to PGH2 (or 2-PGH2-glycerol); and (3) isomerization of unique 18 amino acid insert near its carboxyl terminus that
PGH2 (or 2-PGH2-glycerol) to one of the major prostanoids by regulates the degradation of the protein. The reason for the
a terminal synthase. Mobilization of AA involves cytosolic existence of the two PGHS isoforms is not known. Each protein
phospholipase A2 (cPLA2) which translocates to the endoplas- functions as a homodimer with a monomer mass of 72 kDa.
mic reticulum (ER) and nuclear membrane (NM) when intra- PGHSs are integral membrane proteins which instead of hav-
cellular Ca2þ concentrations rise and cleaves AA from a ing transmembrane helices have a cluster of four amphipathic
phospholipid on the cytosolic surface of the membrane. helices that interact with only one face of the lipid bilayer.
Newly released AA moves, methyl end first, to the luminal PGHSs are inhibited by nonsteroidal anti-inflammatory drugs
half of the bilayer where it enters the active site of a prostaglan- (e.g., aspirin, ibuprofen, and naproxen), which compete with AA
din endoperoxide H synthase (PGHS) and is oxygenated to for binding to the COX site of PGHSs. Prostaglandin synthesis
PGG2 and its 15-hydroperoxyl group is then reduced to form mediated by PGHS-2 can be inhibited by anti-inflammatory
PGH2; PGHSs are also called cyclooxygenases (COXs) and steroids, which block the synthesis of PGHS-2 protein. Drugs
there are two isoforms (PGHS-1 and -2; COX-1 and -2). Once called COX-2 inhibitors (e.g., celecoxib) target PGHS-2 specifi-
formed, PGH2 is isomerized by a synthase located in the ER cally and are used as anti-inflammatory and analgesic agents.

581
582 Lipids Carbohydrates Membranes and Membrane Proteins | Prostaglandins and Leukotrienes

Aspirin, acetylsalicylic acid, binds the COX site of PGHSs and but the PGHS-2 gene is 8 kb and the PGHS-1 gene is 22 kb. Little
acetylates Ser530 causing irreversible inhibition of PGHSs is known about the regulation of PGHS-1 gene expression.
(Figure 2). Low doses of aspirin are used prophylactically to PGHS-2 gene transcription can be induced by cytokines and
reduce coronary thrombosis; at low doses, aspirin inhibits growth factors that function through multiple response ele-
PGHS-1 in platelets blocking thromboxane formation. PGHS ments in the PGHS-2 gene promoter. There is much interest in
inhibitors appear to reduce mortality from colon cancer. this because of the potential role of altered PGHS-2 expression
in carcinogenesis and inflammatory diseases.

PGHS-1 and PGHS-2 Gene Expression

PGH2 Metabolism
There are separate genes for PGHS-1 and PGHS-2 located on dif-
ferent chromosomes. They have similar intron/exon structures Different cells form different prostanoids. PGE2, PGD2, PGF2,
PGI2, and TXA2 syntheses from PGH2 are catalyzed by PGE,
PGD, PGF, PGI, and TXA synthases, respectively; all of these
Phospholipid
enzymes catalyze isomerization reactions. PGF2 formation
Phospholipase A2 involves a two-electron reduction of PGH2. PGI and TXA
synthases are cytochrome P-450s. PGE synthase is a member
COOH
H of the MAPEG family of transmembrane ER proteins and
requires reduced glutathione for activity. There are two PGD
synthases: a hematopoietic type is glutathione dependent and
Arachidonic acid
has been isolated from spleen and a lipocalin type is glutathi-
Cyclooxygenase one independent and occurs in brain.
2O2

COOH
O Prostanoid Catabolism
PGHS
O
The initial step in the inactivation of PGE2 is oxidation
PGG2 OOH
Peroxidase of the 15-hydroxyl group to a 15-keto group catalyzed by
2e- 15-hydroxyprostaglandin dehydrogenase. Further catabolism
COOH involves reduction of the double bond between C-13 and
O C-14, b-oxidation, and o-oxidation.
O
PGH2
OH Prostanoid Actions
Synthases
There are pharmacologically distinct receptors for each of the
known prostanoids. In the case of PGE2, four different prosta-
Prostaglandins Thromboxane A2
(D2, E2, F2a, and l2)
glandin E (EP) receptors have been identified and designated
as EP1, EP2, EP3, and EP4 receptors. EP1 is coupled through Gq
Figure 1 Biosynthesis of common prostanoids from AA. to the activation of phospholipase C, EP2 and EP4 are coupled

HIS207
H
N
Arachidonate
N H Hydroperoxide

O TYR385
O R
C O
H N N
–O
+ Fe3+
NH2 H N N
13 OH

C H
HEME
NH2 N
O
ARG120
CH2 N HIS388

SER530

Figure 2 Model of the cyclooxygenase and peroxidase active sites of the ovine PGHS-1.
 Lipids Carbohydrates Membranes and Membrane Proteins | Prostaglandins and Leukotrienes 583

via Gs to the stimulation of adenylate cyclase, and EP3 receptors whereas mast cells and eosinophils express LTC4 synthase
are coupled via Gi to inhibition of adenylate cyclase. and produce LTC4.
EP3 receptors are involved in the development of fever, EP2
and EP4 function in bone resorption, and EP1 receptors are
5-Lipoxygenase
involved in chemically induced colon cancer. Prostanoids can
activate some isoforms of peroxisomal proliferator-activated
Human 5-LO (E.C.1.13.11.34) is a nonheme iron-containing
receptors (PPARs). PGI2 can be involved in PPARd-mediated
protein of 673 amino acids. It catalyzes the abstraction of a
responses such as decidualization and apoptosis.
hydrogen atom from C-7 of AA and insertion of O2 at C-5 to
generate 5(S)-HpETE (Figure 3). The LTA4 synthase activity of
5-LO then catalyzes removal of 10 pro-R hydrogen atoms from
Leukotrienes and Lipoxygenase Products 5(S)-HpETE through a second redox cycle and an internal
rearrangement of double bonds to form LTA4. LTA4, has a
Leukotrienes were discovered in 1979 during the search for half-life of less than 10 s at pH 7.4. Purified 5-LO requires
the chemical structure of the “slow reacting substance of Ca2þ, ATP, fatty acid hydroperoxide, and phosphatidylcholine
anaphylaxis.” Leukotrienes are produced by the action of in addition to the AA and O2 substrates. Ca2þ facilitates the
5-lipoxygenase (5-LO) (Figure 3) which catalyzes both an association of 5-LO with internal membranes. The need for
oxygen insertion to form 5-HpETE and a dehydration reaction an increase in intracellular Ca2þ concentrations differentiates
to form leukotriene A4 (LTA4). LTA4 is converted to the biolog- 5-LO from other lipoxygenases. 5-LO oxygenates AA at the
ically active leukotrienes LTB4 and LTC4 by LTA4 hydrolase and nuclear member (NM). When Ca2þ concentrations increase
LTC4 synthase, respectively. LTB4 is a potent chemotactic and in neutrophils and mast cells, 5-LO and cytosolic PLA2 become
chemokinetic agent for human polymorphonuclear leuko- associated with the nuclear membrane where another protein
cytes, whereas LTC4 constricts specific smooth muscle (e.g., required for LTA4 synthesis – 5-lipoxygenase activating protein
bronchial smooth muscle) and mediates leakage of vascular (FLAP) – is found. In other cells 5-LO is constitutively asso-
fluid in the process of edema. The formation of LTC4 versus ciated with the NM, whereas in alveolar macrophages 5-LO is
LTB4 is controlled by the expression of LTA4 hydrolase or present in the nucleus. The drug zileuton inhibits 5-LO.
LTC4 synthase by specific cell types. For example, the human The 5-LO gene is large (c. 80 kb) and has 14 exons. It is on
neutrophil expresses LTA4 hydrolase and produces LTB4, human chromosome 10. The human 5-LO gene promoter

Phospholipids

cPLA2

O
OH

HOO O 5-Lipoxygenase

OH Arachidonic acid

5-HpETE HO HO OH
O
5-Lipoxygenase
LTB4
O O LTA4
hydrolase
OH

LTA4 HO OH
LTC4 O
synthase S
CONHCH2COOH
NHCO(CH2)2CHCOOH

LTC4 NH2

Figure 3 Biochemical pathway of the conversion of AA into the biologically active leukotrienes. AA released from phospholipid by cPLA2 is
metabolized by 5-LO to 5-hydroperoxyeicosatetraenoic acid (5-HpETE) and leukotriene A4 (LTA4) which is then converted to LTB4 by LTB4 hydrolase
or conjugated to glutathione to form LTC4 by LTC4 synthase.
584 Lipids Carbohydrates Membranes and Membrane Proteins | Prostaglandins and Leukotrienes

contains Spl and EGR-1-binding sites at –88 to –212 bp can occur from the C-1 carboxyl moiety of LTB4 resulting in
upstream of the translation start site. Genetic polymorphisms the loss of the C-5 hydroxyl group and from the 20-carboxy
in humans are found in Spl-binding sites. terminus of 20-carboxy-LTB4. Individuals with genetic deficien-
cies in peroxisomal metabolism (Zellweger disease) that reduce
5-Lipoxygenase Activating Protein b-oxidation or in aldehyde dehydrogenase (Sjogren–Larsson
syndrome) excrete intact LTB4 and 20-hydroxy-LTB4. LTC4 catab-
FLAP was discovered as a novel protein essential for leukotri- olism involves peptide cleavage reactions involving glutamyl
ene synthesis which was the target of an investigational drug transpeptidase and various dipeptidases to yield LTD4 and
from Merck (MK886). FLAP is an integral membrane protein of LTE4, both of which have biological activity. The sulfur atom of
the NM containing 161 amino acids. The function of FLAP is sulfidopeptide leukotrienes can be oxidized by reactive oxygen
unclear but it does bind AA analogs suggesting that it may species. More specific metabolic processing of the sulfidopeptide
transfer AA to 5-LO. LTC4 synthase has 31% amino acid iden- leukotrienes includes o-oxidation by cytochrome P-450 followed
tity to FLAP with a highly conserved region putatively involved by b-oxidation from the o terminus.
in AA binding.
Leukotriene Biology and Leukotriene Receptors
LTA4 Hydrolase and LTC4 Synthase
LTB4 functions in inflammatory processes through its chemo-
LTA4 hydrolase catalyzes the stereochemical addition of water tactic and chemokinetic affect human polymorphonuclear leu-
to C-12 of LTA4 to form the neutrophil chemotactic factor kocytes. LTB4 induces the adherence of neutrophils to vascular
LTB4. LTA4 hydrolase contains 610 amino acids and one essen- endothelial cells and enhances the migration of neutrophils
tial zinc atom. The enzyme is structurally related to zinc metal- (diapedesis) into extravascular tissues. The biological activity
loproteases and exhibits protease activity; bestatin and of LTB4 is mediated through two specific G-protein-coupled
captopril inhibit the enzyme. LTA4 and LTC4 synthases are receptors termed BLT1 and BLT2. LTC4 and its peptide cleav-
found in many cells including those which do not contain age products LTD4 and LTE4 mediate bronchial smooth
5-LO. It is believed that such cells can cooperate in LTB4 and muscle contraction in asthma and cause edema. There are
LTC4 transcellular synthesis by transfering LTA4 from one cell two G-protein-linked receptors for cysteinyl leukotrienes called
to another. LTA4 hydrolase is localized in the cytosol. To CysLT1 and CysLT2. CysLT1 is found in bronchial and intesti-
metabolize the unstable LTA4, LTA4 hydrolase must either nal smooth muscle; LTD4 and LTC4 activate CysLT1. Montelu-
come in contact with the NM during LTA4 formation or a kast, pranlukast, and zafirlukast inhibit the CysLT1 receptor
carrier protein must transfer LTA4 to LTA4 hydrolase. The crys- in humans. CysLT2 receptors are abundant in the heart and
tal structure of LTA4 hydrolase has been determined. in vascular endothelial cells.
LTC4 synthase (E.C.2.5.1.37) conjugates reduced glutathi-
one (g-glutamyl-cysteinyl glycine) to LTA4. LTC4 synthase is
See also: Bioenergetics: Cytochrome P-450; Signaling:
located on the NM. The enzyme has significant sequence
Eicosanoid Receptors; Phospholipase C.
homology to microsomal glutathione-S-transferases and
FLAP. LTC4 synthase is found predominantly in mast cells,
macrophages, eosinophils, and monocytes. MK-886, which
interacts with FLAP, also inhibits LTC4 synthase. The gene for Further Reading
LTC4 synthase is located on human chromosome 5.
Evans J, Ferguson A, Mosley R, and Hutchinson J (2008) What’s all the FLAP about?:
5-Lipoxygenase-activating protein inhibitors for inflammatory diseases. Trends in
Leukotriene Metabolism Pharmacological Sciences 29: 72–78.
Funk C (2005) Leukotriene modifiers as potential therapeutics for cardiovascular
disease. Nature Reviews Drug Discovery 4: 664–672.
LTB4 can be oxygenated to 20-hydroxy-LTB4 by specific cyto-
Murphy R and Gijon M (2007) Biosynthesis and metabolism of leukotrienes.
chrome P-450s of the CYP4F family. 20-Hydroxy-LTB4 can be Biochemical Journal 405: 379–395.
further metabolized to 20-carboxy-LTB4 by CYP4F3 or to Rouzer CA and Marnett LJ (2009) Cyclooxygenases: Structural and functional insights.
20-oxo-LTB4 by alcohol dehydrogenase then to 20-carboxy-LTB4 Journal of Lipid Research 50: S29–S34.
by fatty aldehyde dehydrogenase. An alternative pathway in- Smith WL (2008) Nutritionally essential fatty acids and biologically indispensable
cyclooxygenases. Trends in Biochemical Sciences 33: 27–37.
volves an initial oxidation of the 12-hydroxy group to a 12-oxo Smith WL and Murphy RC (2008) The eicosanoids: Cyclooxygenase, lipoxygenase, and
moiety followed by reduction of the conjugated dienone and the epoxygenase pathways. In: Vance DE and Vance JE (eds.) Biochemistry of Lipids,
10,11 double bond. A secondary catabolic pathway, b-oxidation, Lipoproteins and Membranes, 5th edn., pp. 331–362. Amsterdam: Elsevier.