Hydroxyeicosatetraenoic Acids and Related
The oxygenated metabolites or oxylipins derived from arachidonic and related fatty acids are produced through a series of complex interrelated biosynthetic pathways often termed the 'eicosanoid cascade'. Here, the linear hydroxyeicosatetraenes and related mono-oxygenated metabolites are described, together with octadecanoids produced from linoleate and similar oxylipins from eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. While these are relatively simple in structure, they are precursors for families of more complex molecules, such as the leukotrienes and lipoxins and the protectins, resolvins and maresins or 'specialized pro-resolving mediators'. The two main enzymatic pathways for production of these eicosanoid utilize lipoxygenases (LOXs) and oxidases of the cytochrome P-450 family as described below, although some are produced also as a minor byproduct of cyclooxygenases or when cyclooxygenase 2 is inhibited by aspirin, as discussed in our web page on prostanoids, which have distinctive ring structures in the centre of the molecule and are discussed on their own web page. Hydroperoxides can also be formed non-enzymatically as discussed in our web page dealing with isoprostanes.
1. Lipoxygenases and Hydroxyeicosatetraenoic Acids
Lipoxygenases are a family of enzymes that can be characterized as non-heme iron proteins or dioxygenases, which catalyse the abstraction of hydrogen atoms from bis-allylic positions (1Z,4Z-pentadiene groups) of polyunsaturated fatty acids followed by stereospecific addition of dioxygen to generate hydroperoxides. They occur widely in plants, fungi, a few prokaryotes (cyanobacteria and proteobacteria) and animals, but not in the archaea and perhaps insects. The plant lipoxygenases have distinctive substrates and products, and they are described in our web page dealing with plant oxylipins rather than here, although interesting parallels can be drawn with the mechanisms and functions of the animal enzymes.
Animal lipoxygenases that utilize arachidonic acid as substrate are of great biological and medical relevance, because of the functions of the products in signalling or in inducing structural or metabolic changes in the cell. For example, they react with arachidonic acid per se to produce specific hydroperoxides and thence by downstream processing the plethora of eicosanoids, each with distinctive functions, which are described in this and other web pages and include lipoxins, trioxilins, leukotrienes, hepoxilins, eoxins and specialized pro-resolving mediators. Only the primary lipoxygenase products are discussed in this web page. However, these enzymes can also react directly with phospholipids in membranes to produce hydroperoxides that perturb the membrane structure. Thence, programmed structural changes in the cell can be induced, as in the maturation of red blood cells. Beyond their role in oxylipin production, lipoxygenases and lipid hydroperoxides can stimulate the formation of secondary products, and for example, lipoxygenases can attack low-density lipoproteins directly with major implications for the onset of atherosclerosis. They have a more general function in cellular redox homeostasis.
The nomenclature of animal lipoxygenases is based on the specificity of the enzymes with respect to the products of the reaction with arachidonate (not the initial point of hydrogen abstraction); for example, 12-LOX oxygenates arachidonic acid at carbon-12. The stereochemistry of the reaction can be specified when necessary (e.g. 12R-LOX or 12S-LOX), although the more important enzymic hydroperoxides have the S‑configuration. Where more than one enzyme has the same specificity, it may be named after the tissue in which it is found, and there are platelet, leukocyte and epidermal types of 12‑LOX, for example.
As the research in this area has developed, this simplistic nomenclature has become confusing. It has become evident that some enzymes can oxygenate more than one position and that this can vary with the chain-length of the polyunsaturated substrate and the positions of the double bonds. Enzymes with specificities for four different positions in arachidonic acid occur in animal tissues, i.e. 5-LOX, 8-LOX, 12 LOX, and 15-LOX, although some of these have dual specificities, while many iso-forms exist depending on species. There are now considered to be six main lipoxygenase family members in humans (5‑LOX, 12‑LOX, 12/15‑LOX (15‑LOX type 1), 15‑LOX type 2, 12(R)-LOX, and epidermal LOX (eLOX-3) and seven in mice. Orthologues of the same gene have different reaction specificities in different species. For example, mice do not express a distinct 15-LOX but rather a leukocyte-derived 12-LOX with some 15‑LOX activity, so it can be difficult to extrapolate from animal experiments to human conditions; the human enzymes only are discussed at length below. The positions at which the enzymes interact with arachidonic acid and the main products are illustrated in the figure below.
Each of the lipoxygenase proteins in animal tissues has a single polypeptide chain with a molecular mass of 75-80 kDa. They have a N‑terminal 'β‑barrel' or 'PLAT' domain, which is believed to function in the acquisition of the substrate, and a larger α-helical catalytic domain containing a single atom of non-heme iron, which is bound to four conserved histidine residues and to the carboxyl group of a conserved isoleucine at the C-terminus of the protein. The PLAT domain anchors the otherwise cytosolic protein to membranes in response to intracellular calcium levels. For catalysis, the iron component of the enzymes must be oxidized to the active ferric state.
All of the enzymes appear to include the fatty acid substrate within a tight channel with smaller channels that direct molecular oxygen toward the selected carbon, facilitating the formation of specific hydroperoxy-eicosatetraenes (HPETEs). In other words, the regiospecificity is regulated by the orientation and depth of substrate entry into the active site, while stereospecificity is controlled by switching the position of oxygenation on the reacting pentadiene of the substrate at a single active enzyme site, which is conserved as an alanine residue in S‑lipoxygenases and a glycine residue in the rarer R‑lipoxygenases. There is evidence that two amino acids opposite the catalytic iron ion determine the orientation of the substrate for entry into the enzyme channel. With 5-LOX and 8-LOX, the carboxyl group of arachidonic acid enters the active site first, while with 12-LOX and 15-LOX the ω-terminus enters the site and facilitates the activity. It should be noted that the specificities of the enzymes are not always absolute and can differ between species. The N-terminal domains function in membrane binding and regulation, and are not required for the catalytic activity.
Lipoxygenase action is believed to proceed in four steps - hydrogen abstraction (1), radical rearrangement (2), oxygen insertion (3), and peroxy radical reduction (4), all occurring under strict steric control, as illustrated.
For example, in the action of 5-LOX, the first and rate-limiting step is the abstraction of a hydrogen atom from carbon 7 of arachidonic acid by non-heme ferric iron (Fe(III)), involving a proton-coupled electron transfer in which the electron is transferred directly to the iron and the proton is acquired simultaneously by the hydroxide ligand in a concerted mechanism to produce a substrate radical, while the iron atom is reduced to the ferrous form (Fe(II)). The cis-double bond in position 5 migrates to position 6 to form a more stable conjugated diene with a change to the trans-configuration, before dioxygen is introduced opposite to the removed hydrogen (antarafacially) to generate a lipid peroxyl radical. Finally, the lipid peroxyl radical is reduced by Fe(II) and protonated to form a lipid hydroperoxide in another concerted reaction. In the process, the iron atom is re-oxidized to its ferric form for another round of catalysis. In this example, the resulting product is 5S‑hydroperoxy-6t,8c,11c,14c-eicosatetraenoic acid (5‑HPETE).
HPETE in general have a short half-life and are rapidly metabolized to hydroxy-eicosatetraenes (HETE) with the same stereochemistry, often via reduction by the abundant and ubiquitous glutathione peroxidases (step 5). While their primary function is to act as intermediates in the biosynthesis of other eicosanoids, HPETE have some biological activities of their own.
Alternatively, isomerization reactions can occur to produce leukotrienes and lipoxins via epoxy intermediates. Simplistically, the Fe2+in the lipoxygenase cleaves the O-O bond in the hydroperoxide with transer of the hydroxyl group to form Fe3+-OH. The residual alkoxyl radical is cyclized to form an epoxy fatty acid.
5-LOX (ALOX5) is found only in cells derived from bone marrow (leukocytes, macrophages, etc) and it is of particular interest as the product is the primary precursor for the leukotrienes and lipoxins and for resolvins. It is a cytosolic protein when intracellular calcium levels are low and becomes associated with the nuclear membrane when they are high or after phosphorylation. In contrast to other lipoxygenases, it requires the presence of a specific activator protein, lipoxygenase-activating protein (FLAP), on the perinuclear membrane. This facilitates the transfer of arachidonic acid to the active site on 5-LOX and is believed to effect the functional coupling of phospholipase A2 (cPLA2) to 5-LOX at the membrane. It is noteworthy that both cPLA2 and 5-LOX are Ca2+-dependent. The activities of 5-LOX and related enzymes are regulated by a number of factors that include the concentration and availability of the substrates, the redox state, intracellular Ca2+ concentrations and phosphorylation-dephosphorylation by means of various protein kinases.
8-, 12- and 15-LOX operate in a similar way to give analogous products and associate with membranes in a calcium-dependent process, although they do not require accessory proteins. 15‑LOX exists in two forms, but the more important has a broader specificity (not recognized in its name) and is expressed primarily in reticulocytes and macrophages on stimulation by interleukins 4 and 13. This form is sometimes termed 15‑LOX‑1 (or ALOX15 or 12/15‑LOX) as it can also produce some 12‑HETE, 8,15‑diHETE and eoxin A4 from arachidonic acid. In addition, it is able to oxidize linoleate to 13‑hydroperoxy-octadecadienoate (and in part to the 9-isomer), as well as oxidizing α-linolenic, γ-linolenic, eicosapentaenoic and docosahexaenoic (DHA) acids. For example, it reacts with DHA to produce 17(S)-HPDHA, a precursor of resolvins and protectins. 15‑LOX‑1 is induced by the action of cytokines, and uniquely, it synthesises both pro- and anti-inflammatory molecules. Molecular genetics studies show that this broad reactivity of the enzyme is seen only in higher ranked primates and not in mammals ranked in evolution lower than gibbons, where the enzyme has 12‑lipoxygenating specificity with arachidonate.
The second human arachidonate 15‑lipoxygenase has 40% homology with the first and is termed 15‑LOX‑2 (or ALOX15B). The human form of 15‑LOX‑2 produces 15-HETE exclusively and is expressed constitutively in macrophages (although the function of the enzyme is not clear) and also in prostate, lung, skin and cornea. Further, both forms of 15-LOX differ from the other lipoxygenases in that they can utilize these fatty acids bound to intact lipids, including phospholipids and cholesterol esters in biomembranes and lipoproteins, as substrates. Hence the interest in the role of the enzyme in autophagy, membrane disruption and in disease states. Mouse skin produces a lipoxygenase (8-LOX) that is structurally related to 15-LOX-2, but generates 8S-HETE and 8S,15S-diHPETE from arachidonic acid. Some 15(R)-HETE is produced by the action of COX-2 and aspirin.
12(S)-LOX (ALOX12) from human platelets and leukocytes was one of the first lipoxygenases to be characterized, but a rather different enzyme is present in the epidermis. Although lipoxygenase metabolites generally have a hydroperoxide moiety in the S-configuration, lipoxygenases in mammalian skin can produce the R‑form. Indeed, 12R‑HETE was first characterized as a component of psoriatic lesions. One of the enzymes responsible is a second form of the human 15‑lipoxygenase (15-LOX-2), but there is also a 12R-LOX (ALOX12B) with quite specific functions in keratinocytes and certain other tissues, especially in relation to linoleate metabolism and the formation of essential ceramides in the corneocyte envelope. Thus, eLOX3 exhibits a hydroperoxide isomerase activity (lipohydroperoxidase activity), and transforms hydroperoxides to epoxy-alcohols and ketones. Enzymes related to the last are common in aquatic invertebrates.
Hydroxy fatty acids produced by lipoxygenases can be further oxidized to their keto analogues (c.f. 5-oxo-eicosatetraenoic acid below) or to dihydroxy derivatives that include the leukotrienes discussed in a separate web page. In addition, some have been reported to form glutathione conjugates.
Fungal and bacterial enzymes: Certain fungi and yeasts are able to produce 3R- and/or 3S-HETE and 3,18-di-HETE, when supplied with exogenous arachidonic acid from their animal hosts during infection. The biochemical mechanism is unclear, and there are reports that implicate lipoxygenases, cyclooxygenases and mitochondrial β-oxidation. With some pathogenic fungi, the 3-hydroxyeicosanoids produced in infected cells can be acted upon by the host COX-2 enzyme to form a family of 3-hydroxy-prostaglandins, which are at least as active biologically as the normal compounds. Aspergillus fumigatus has two lipoxygenase homologues of human ALOX15 and ALOX5, termed LoxA and LoxB, of which the latter is secreted extracellularly and produces 13‑hydroxyoctadecadienoic acid (13-HODE) from linoleate. Prokaryotic LOX-encoding genes are known from Pseudomonas aeruginosa (Gram-negative bacteria) with the capacity to produce 15S‑hydroxyeicosatetraenoic acid (15S‑HETE), but these are distant evolutionarily from human LOX.
2. Cytochrome P450 Oxidases and Hydroxy-/Epoxyeicosatetraenoic Acids
Arachidonic acid can be oxidized by several cytochrome P450 mixed-function oxidases to produce various HETE isomers (the name was coined to describe the first such enzyme to be characterized and was based on an unusual absorbance peak at 450 nm from its carbon monoxide-bound form). These enzymes are a superfamily of membrane-bound hemoproteins that catalyse the scission of the dioxygen bond in molecular oxygen and transfer of a single atomic oxygen to a substrate carbon atom, i.e. they are monooxygenases (with the release of the other oxygen atom as water). The result is the introduction of either a hydroxyl or an epoxyl group into the molecule. The catalytic turnover of the reaction is NADPH-dependent, requiring transfer of electrons from NADPH to the P450 heme iron (lipoxygenases use non-heme iron), for which a membrane-bound enzyme partner, NADPH-cytochrome P450 reductase, is essential in the endoplasmic reticulum (or functionally related enzymes in mitochondria).
Cytochrome P450 oxidases are found in all mammalian cell types and indeed appear to be ubiquitous in all living organisms, although the number and distribution of particular forms of the enzymes are specific both to cell type and species. They are located in the endoplasmic reticulum with a limited expression in mitochondria (and perhaps plasma membrane and nucleus), and predominantly in the liver but with significant levels in some other extrahepatic tissues, including brain, kidney and lung. In addition to their role in generating HETE isomers, enzymes of this kind have a more general function in the eicosanoid cascade in the metabolism of prostanoids, and they are involved in cholesterol and steroid metabolism as well as detoxification of lipophilic xenobiotics, including drugs and chemical carcinogens. Their nomenclature starts with the root 'CYP', followed by a number allocated to the family, a letter for subfamily, and a gene-identifying number for isoforms.
CYPs have two domains: a β-sheet-rich N-terminal domain and a larger helix-rich C-terminal catalytic domain. Those enzymes in the endoplasmic reticulum have a transmembrane helix in the N-terminal domain that is required for membrane anchoring, but this feature is not present in mitochondrial CYPs, which rely upon hydrophobic regions on the surface to bind to membranes. The catalytic domain contains the heme prosthetic group in a deep cavity, where variability in the structure of the active site in each form explains the flexibility for substrates and products. Access channels permit entry of substrates and exit channels allow egress of the product.
Three types of reaction have been observed in animal cells, leading to the formation of three distinct families of eicosanoids, all requiring unesterified arachidonic acid as substrate, although appreciable amounts of the products can be found in esterified form (see below). The CYP1 to 3 families tend to catalyse epoxidation and hydroxylation reactions, while the CYP4 family favours hydroxylation and especially ω-hydroxylation. As an example, one series of reactions occurs at bis‑allylic centres and is lipoxygenase-like in the nature of the ultimate HETE products, although hydroperoxy intermediates are not involved. Thus, microsomal cytochrome P450 oxidases can react with arachidonic acid to produce six regioisomeric cis,trans-conjugated dienols, i.e. with the hydroxyl group in positions 5, 8, 9, 11, 12 or 15. The mechanism is believed to involve bis-allylic oxidations at either carbon-7, 10 or 13, followed by acid-catalysed rearrangement to the cis,trans-dienol. Two examples of the products are illustrated. 12(R)-HETE as opposed to the 12(S)-isomer is the main product of the reaction, and this was at one time though to be a distinguishing feature, but some other lipoxygenases are now know to produce the former enantiomer.
Secondly, there are ω- and (ω-1)-hydroxylases that introduce a hydroxyl group into positions 20 and 19, respectively, of arachidonic acid mainly, although other enzymes can react at positions 16, 17 and 18 also. The reaction was first observed with medium-chain saturated fatty acids, such as lauric (12:0), where it may play a role in oxidative catabolism. Some isoenzymes are specific for laurate, others for arachidonate, and some will utilize both fatty acids as substrates. In humans, the iso-forms CYP4A and CYP4F are the main enzymes involved in ω‑hydroxylation of polyunsaturated fatty acids, including both arachidonic and eicosapentaenoic acids, while the CYP1A1, CYP2C19, and CYP2E1 forms effect (ω‑1)-hydroxylation. Both R- and S-forms of the sub-terminal HETE can be produced with differing biological activities.
20-HETE is metabolized by cyclooxygenases into a hydroxy analogue of prostaglandin H2 (20-OH PGH2), a vasoconstrictor that is further converted by isomerases into 20-OH PGE2 and 20-OH PGI2 (vasodilator/diuretic metabolites) and 20‑OH thromboxane A2 and 20-OH PGF2α (vasoconstrictor-antidiuretic metabolites). Other CYP450 enzymes are capable of introducing hydroxyl groups in positions 2 and 3 of fatty acids, while others can effect decarboxylation or form terminal alkenes, properties that have biotechnology potential.
Epoxyeicosatrienoic acids: The third series of reactions of P450 arachidonic acid monooxygenases involves the formation of epoxytrienoic acids (‘EET’) from arachidonic acid, i.e. four cis-epoxyeicosatrienoic acids (14,15-, 11,12-, 8,9-, and 5,6-EETs). Apart from the 5,6-isomer, they are relatively stable molecules.
Several iso-enzymes of the cytochrome P450 epoxygenase exist, with CYP2C and CYP2J as the most active, and they can produce all four EET regioisomers, although one isomer usually tends to predominate in a given tissue. For example, epoxygenases that produce 14,15-EET as the main isomer also synthesise a significant amount of 11,12‑EET and a little 8,9-EET. The epoxygenase attaches an oxygen atom to one of the carbons of a double bond of arachidonic acid, and as the epoxide forms the double bond is reduced. The enzymes are located both in the cytosol and the endoplasmic reticulum of endothelial cells, and they make use of arachidonic acid that is hydrolysed from phospholipids when the Ca2+-dependent phospholipase A2 is activated and translocated from the cytosol to intracellular membranes.
The proportions of the various isomers depend on tissue and species, although the 11,12- and 14,15‑EET generally tend to predominate. In the rat, 14,15‑EET amounts to about 40% of those produced in the heart, while 11,12-EET represents 60% of those produced in the kidney, for example. To add to the complication, each of these regioisomers is a mixture of R,S- and S,R-enantiomers, and each iso-enzyme produces variable proportions, differing even among regioisomers. Eight isomers can be formed, therefore, each with somewhat different biological activities. Adrenic acid (22:4(n-6)) is converted similarly to epoxy metabolites, and 16,17‑epoxydocosatrienoic acid is the most abundant isomer, especially in liver.
The epoxygenases require the fatty acid substrate to be in the unesterified form, but the products can be esterified later. Thus, significant amounts of epoxyeicosatrienes are found esterified to position sn-2 of phospholipids, including phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol, perhaps as a storage form that is available when a rapid response is required. For example, free epoxyeicosatrienes are released following activation of phospholipase A2 by neuronal, hormonal or chemical stimuli. There is also a possibility that esterified epoxy-eicosanoids may have a biological function within membranes. The presence of esterified EETs in plasma, suggests that some exchange between tissues is possible, although most are believed to be produced close to the site of action. In many tissues, the esterified epoxy-eicosanoids are so similar in composition to those in the free form, that the conclusion must be that they are entirely products of enzyme action. On the other hand, non-enzymic lipid peroxidation has been observed in erythrocytes in vitro, and some EETs with the epoxide group in both the cis- and trans-configurations may arise by this route.
EETs are rapidly metabolized in vivo to the corresponding dihydroxyeicosatrienoic acids (DHET) by epoxide hydrolases, of which at least five forms are known with different cellular locations and preferred substrates. The cytosolic (EPHX2) and membrane-bound (EPHX1) enzymes are of special importance, both in terms of lipoxin metabolism and for detoxification of xenobiotic epoxides. EPHX2 is widely expressed throughout the body, and in humans, this is a 62kDa enzyme composed of two domains separated by a short proline-rich linker in which the N-terminal domain has phosphatase activity towards lipid phosphates, while the C-terminal domain has the epoxide hydrolase activity. The reaction is illustrated below for the conversion of 14,15-EET to 14,15-DHET.
This enzyme metabolizes 8,9-, 11,12- and 14,15-EET efficiently, but 5,6-EET is a poor substrate. It also displays some enantioselectivity and this may be an important factor in determining the stereochemistry of the circulating epoxides. In addition, 11,12- and 14,15-EET can undergo partial β-oxidation to form C16 epoxy-fatty acids, or they can be elongated to C22 products. 5,6- and 8,9-EET are substrates for cyclooxygenase. While DHETs were once believed to be merely deactivation products of EETs, they are now known to have some biological effects in their own right.
3. 5-Oxo-Eicosatetraenoic Acid
5-Oxo-6t,8c,11c,14c-eicosatetraenoic acid (5-Oxo-ETE) is a metabolite of 5S-hydroxy-6t,8c,11c,14c-eicosatetraenoic acid (5‑HETE), produced by oxidation by NADP+-dependent 5-hydroxy-eicosanoid dehydrogenase, an enzyme found in the microsomal membranes of white blood cells (leukocytes), platelets and especially of eosinophils and neutrophils. The enzyme requires the presence of a 5S‑hydroxyl group and a trans-6 double bond in the eicosanoid, and NADP+ is a cofactor. Synthesis of the metabolite is stimulated during periods of oxidative stress. In addition, some 5-oxo-ETE may be formed directly from 5‑hydroperoxyeicosatetraenoic acid, possibly by a non-enzymic route. In neutrophils, a high proportion is rapidly incorporated into the triacylglycerol fraction.
It appears that 5-hydroxyeicosanoid dehydrogenase can also catalyse the reverse reaction, i.e. the reduction of 5-oxo-ETE, and this seems to be of particular important in platelets. The biological activity of 5-oxo-ETE is of course changed by this reverse reaction, and alternative deactivation can occur by reduction of the double bond in position 6, or by further oxidation either by lipoxygenases or by cytochrome P450 enzymes, the latter in positions 19 or 20.
In fact, all the HETE isomers can be converted to oxo-metabolites by specific hydroxy-eicosanoid dehydrogenases, and the 11-, 12- and 15‑isomers possess appreciable biological activity. For example, 15(S)-HETE and 11(R)-HETE are substrates for 15-hydroxyprostaglandin dehydrogenase, the enzyme involved in the first step of prostaglandin catabolism, to yield 15-oxo-ETE and 11-oxo-ETE, respectively, which mediate anti-proliferative properties in endothelial cells. Similarly, 14-hydroxy-docosahexaenoic acid is an good substrate for the enzyme to yield the 14-oxo analogue. It is now recognized that α,β-unsaturated keto-eicosanoids generated in this way are electrophilic and have the potential to interact with nucleophilic centres in proteins and other molecules to modify their activities. However, 11- and 15‑oxo‑ETE form CoA esters which can undergo up to four double bond reductions.
4. Mono-oxygenated Metabolites of EPA and DHA
Lipoxygenases and cytochrome P450 oxidases interact with the other essential polyunsaturated fatty acids of the omega-3 and omega-6 families, especially the former, to give comparable series of metabolites. For example, lipoxygenases have much the same positional specificity with eicosapentaenoic acid (EPA or 20:5(n-3)) as with arachidonic acid to produce hydroxy-eicosapentaenoic acids (HEPE), i.e. 5- and 12‑HEPE. 18‑HEPE is produced by aspirin-acetylated COX-2 or by CYP2C8/CYP2J2. 5-Lipoxygenase generates 4- and 7-hydroxy metabolites from docosahexaenoic acid (22:6(n‑3) or DHA), while 12-lipoxygenase generates 11- and 14-hydroxy metabolites, and 15‑lipoxygenase (15-LOX-2) produces a 17‑hydroxy metabolite. Further reactions produce the protectins, resolvins and maresins or 'specialized pro-resolving mediators', which have special importance in the resolution of inflammation and have their own web page
The products of the lipoxygenases with arachidonate were soon documented, but it has taken longer to recognize the importance of the metabolites of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, especially the epoxides, produced by the activities of various cytochrome P450 enzymes. Indeed, it is now evident that these n-3 polyunsaturated fatty acids, rather than arachidonic acid, are the preferred substrates for some of the isoforms, specifically the CYP1A, CYP2C, CYP2J and CYP2E subfamily members, which then exhibit very different regio- and stereo-specificities. For example, human CYP1A1 acts mainly as a subterminal hydroxylase with arachidonate producing four different isomers, but with EPA it generates mainly 17(R),18(S)-epoxy-eicosatetraenoate with almost absolute regio- and stereo-selectivity. Similarly, with DHA it epoxidizes the n-3 double bond and produces 19,20‑epoxydocosapentaenoate. A number of other isoforms of the cytochrome P450 enzymes produce epoxides by reaction with an n‑3 double bond in the same manner, some much more rapidly than with arachidonate as substrate. One exception is CYP2C9, which oxidizes EPA to 14,15-epoxy-ETE mainly and DHA to 10,11‑epoxy‑DPE.
The CYP4A/CYP4F subfamilies are the main enzymes that produce 20-hydroxy-eicosatetraenoic acid from arachidonate in mammals, and they hydroxylate the terminal methyl group in EPA and DHA also at the same rate. In addition, human endothelial cells with upregulated COX-2 and treated with aspirin converted EPA to 18R-hydroxyeicosapentaenoic acid with anti-inflammatory properties. 4-Oxo-DHA is present in plasma of rats fed DHA, and has potent anti-tumour effects against breast cancer, although details of its fine structure and biosynthesis are awaited.
By competing with arachidonate, EPA and DHA may modify the action of the various HETE metabolites, but the oxygenated EPA and DHA compounds have biological properties of their own. For example, significant amounts of DHA epoxides, especially 7,8‑epoxydocosapentaenoic acid, are present in the central nervous system of rats, where they ameliorate the effects of inflammatory pain. 17,18-Epoxyeicosatetraenoic acid generated in the gut is an anti-allergic molecule. It has been suggested that such EPA and DHA metabolites may be responsible for some of the beneficial effects associated with dietary n-3 fatty acid intake.
The action of lipoxygenases upon linoleic acid in plant tissues is discussed in the web page on plant oxylipins, but this fatty acid is acted upon by lipoxygenases in animal tissues in a similar way to produce 9- and 13-hydroperoxy- and thence hydroxy-octadecadienoic acids (octadecanoids or 'HODEs') of defined stereochemistry. In particular, 13(S)‑hydroperoxy-9Z,11E-octadecadienoic acid (13S-HPODE) is generated by the action of 15‑lipoxygenase (15-LOX-1) on linoleic acid, and this is reduced to the hydroxy compound, or oxo-octadecadienoic acids, epoxy-octadecenoic acids and epoxy-keto-octadecenoic acids can be formed in further reactions.
12R-LOX reacts readily with linoleate (9,12-18:2) to produce 9R-HPODE. As linoleic acid is a major unsaturated fatty acid in animal tissues, appreciable amounts of these hydroxy and hydroperoxy metabolites can accumulate and influence inflammatory diseases. Indeed, linoleate metabolites are by far the most abundant oxygenated fatty acids in both free and esterified form in human plasma, and in the brain of rat pups. Epoxy-octadeca-monoenoic acids are produced also by insects, where they are believed to be involved in the resolution of cellular and humoral immune reactions.
A further interesting observation is that one of the unique ceramides of skin, O‑linoleoyl-ω-hydroxyacyl-sphingosine, is a substrate for 12R‑LOX with 9R‑hydroperoxy-linoleoyl-ω-hydroxyceramide as the product. This in turn can be converted to hepoxilin-like compounds, i.e. with an epoxyl group, by an enzyme epidermal lipoxygenase 3 (eLOX-3), while trihydroxy compounds. e.g. octadec-9R,10S,13R-trihydroxy-11E-enoate (tri-HOMEs) may be formed subsequently by the action of an epoxide hydrolase, such as the human soluble enzyme. For example, 9R,10S,13R-trihydroxy-11E-octadecenoate is an important oxylipin formed in porcine and human epidermis, where it interacts with the ceramides to aid formation of the waterproof barrier. Tri-HOMEs are also produced in the lung where biosynthesis is believed to involve formation of a 13S‑hydroperoxide by the action of 15-lipoxygenase and proceeds via an epoxide intermediate. Autoxidation occurs also with linoleate to produce the same types of compound but with more variable stereochemistry.
Enzymes of the cytochrome P450 family make a further contribution and linoleic acid is especially a substrate for CYP epoxygenases, CYP2C9 in human liver especially, yielding the linoleic epoxides 9,10- and 12,13‑epoxyoctadecenoic acids, which are sometimes termed leukotoxins (although this name has been applied also to very different microbial metabolites). They are further metabolized by epoxide hydrolase to form the 9,10- and 12,13‑diols, respectively. While they were first found in patients with burns and inflammatory diseases, 12,13-dihydroxy-9Z-octadecenoate (12,13-diHOME) has some beneficial properties (see below). They are detoxified by conversion to the glucuronides. While similar reactions occur with both α- and γ-linolenate in vitro, the biological significance of these metabolites in vivo is not known.
6. Esterified Oxylipins
Most eicosanoid-generating enzymes require free fatty acids as the substrate, and they are unable to oxidize intact phospholipids, although 15‑LOX in human monocytes is an exception (as is murine 12/15-LOX). However, free 5-, 12- and 15-HETEs can be esterified to phospholipids in tissues, often with some specificity, and it has been established that all mammalian long-chain acyl-CoA synthetase isoforms have the capacity to activate HETE for further esterification through the action of membrane-bound O-acyltransferases (thromboxanes may be an exception). The mechanisms for these esterification processes are discussed in our web page on oxidized phospholipids It is evident that many of these reactions depend on specific cell types and lipids, and that cell compartmentalization is an important factor. Non-enzymatic oxidation (autoxidation) of polyunsaturated fatty acids in esterified form occurs by the initial steps described in our web page on isoprostanes.
Eicosanoids of exogenous origin and those generated endogenously appear to be sensed differently. For example, 15-HETE is selectively esterified to phosphatidylinositol in lung and kidney epithelial cells and in aortic endothelial cells, while 12-HETE occurs predominantly in phosphatidylcholine in microsomal membranes. In neutrophils, 5-HETE is incorporated mainly into phosphatidylethanolamine plasmalogens and phosphatidylcholine, while three 12‑hydroxyeicosatetraenoic acid phosphoinositides have been detected in thrombin-activated platelets. More than 90% of the EETs in most plasma and organ tissues are esterified to position sn-2 of glycerophospholipids, and in aortic endothelial cells, 20‑HETE is present in esterified form in several phospholipid classes, Hydroxydocosahexaenoic acids and hydroxyoctadecadienoic acids are also found in esterified form.
Many of these esterified lipoxygenase and oxidase products of phospholipids remain within the membranes where they are believed to serve as a storage form to be released on appropriate stimulation, possibly into other cellular compartments with different biological roles from their unesterified equivalents. On the other hand, such oxidized lipids have the potential to perturb the membrane structure and effect secondary oxygenations that could induce unwanted changes in cells, such as ferroptosis (see our web page on oxidized phospholipids) and their biological activities are discussed further below. Oxidation of low-density lipoprotein by this means may be important for the initiation of atherosclerosis. There are suggestions that cholesteryl arachidonate in the lipoprotein LDL is a good substrate for 15‑LOX‑1 or oxidizing agents derived from it and that the products are a causative factor in this disease (see our web page on cholesterol).
In addition, phospholipids containing EET are substrates for the production of lipid mediators such as 2-epoxyeicosatrienoyl-sn-glycerols, analogous to the endocannabinoid 2-arachidonoylglycerol (and discussed further under this topic). Kidney and spleen, for example, synthesise 2-glycerol derivatives containing 11,12-EET or 14,15-EET, which are endocannabinoids and exert biological effects by activating the CB1 and CB2 receptors. Similarly, phospholipids containing EET are probable substrates for synthesis of EET-ethanolamide in the liver and kidney. Endocannabinoids such as anandamide and synaptamide can be converted directly to various oxygenated derivatives, which can have higher biological activities than their precursors.
7. Biological Activity
A large number of hydroxyeicosatetraenoic acids and related compounds have now been discovered and most of these have some form of biological activity, in vitro at least, and primarily in signalling. In particular, they modulate ion transport, vascular tone, renal and pulmonary functions, and growth and inflammatory responses through both receptor and non-receptor mechanisms. They are released by the action of growth factors and cytokines, and they attain physiological concentrations much higher than those of prostanoids. This is a field that is developing rapidly and it is evident that the picture is complex and very far from complete. A given eicosanoid of this type can have differing functions in different cell types, and its activity may be opposed or modified by another eicosanoid; the balance between them in a cell may be critical. As animal models can have very different isoforms of enzymes, it is often difficult to translate experiments with other species to human conditions. It is not possible to give a comprehensive picture of these manifold biological activities here, as this would require a substantial tome, and only a few of the more important are described briefly below.
HETEs: 5S-Hydroxy-6t,8c,11c,14c-eicosatetraenoic acid (5(S)-HETE) is important as the precursor of the leukotrienes and lipoxins, but it has some biological functions in its own right, although these can be difficult to disentangle from those of its metabolites, which are more active. For example, like its metabolite 5-oxo-HETE, 5(S)-HETE activates neutrophils and monocytes. It is also known to stimulate proliferation of cancer cells in a similar manner to certain leukotrienes, and increased amounts are formed in brain tumours, for example. 5-LOX inhibitors have preventive effects.
5-Oxo-6t,8c,11c,14c-eicosatetraenoic acid is a chemo-attractant for eosinophils and neutrophils and has many functions in such cells, including actin polymerization, calcium mobilization, integrin expression and degranulation. Its signalling functions are mediated via a specific receptor ('OXE'), leading to increased intracellular calcium concentrations and inhibition of cAMP production. Thereby, it stimulates the proliferation of prostate tumour cells. In addition, it is believed to be an important mediator in asthma and other allergic diseases, and efforts are underway to find inhibitors of the specific receptor that may have clinical utility.
Arachidonate 8(S)-lipoxygenase and its product 8S-hydroxy-5c,9t,11c14c-eicosatetraenoic acid (8S-HETE) has only been found in the skin of mice. It is a potent activator of the peroxisome proliferator-activated receptor PPARα, it is an anti-tumorigenic agent towards skin cancer, and it promotes wound healing in the cornea. In contrast, a human orthologue of this enzyme (15-LOX-2) is found in skin, sebaceous glands and prostate tissue but produces 15S-HETE.
12S-Hydroxy-5c,8c,10t,14c-eicosatetraenoic acid (12S-HETE) is the precursor of the hepoxilins but has important functions of its own. In nervous tissue, it modulates membrane properties and stimulates melatonin synthesis, for example. Together with 15(S)‑HETE, it serves as a secondary messenger in synaptic transmission and is involved in learning and memory processes; increased levels are found in Alzheimer's disease. In leukocytes, it promotes chemotaxis and induces the synthesis of heat-shock protein. It can either stimulate or inhibit aggregation in platelets, depending on species and circumstances. For example, 12S‑HETE can cause constriction of blood vessels, but it can inhibit thromboxane (TxA2)-induced platelet aggregation and it stimulates lipoxin synthesis. The serum level of 12-HETE has been shown to be elevated in individuals with coronary heart disease. Particular attention has been devoted to the effects of 12S‑HETE on inhibiting the adhesion of cancer cells to endothelial cells, an activity that is linked to metastasis in cancer of the prostate and is mediated via cell surface signalling and activation of protein kinase C. It also promotes the proliferation of ovarian cancer cells.
The enantiomeric compound 12R-HETE is believed to be involved in the pathophysiology of psoriasis and similar skin diseases, but it is also essential for the development of normal skin. 12R-HETE produced by cytochrome P450 enzymes may have a function in the eye.
12S-Hydroxy-8,10,14-eicosatrienoic acid (12S-HETrE), derived from dihomo-γ-linolenic acid (20:3(n-6)) has been shown to provide protection against thrombotic-mediated events in vivo and has a potential therapeutic role in providing cardioprotection by activating the prostacyclin receptor (IP).
The hydroperoxide precursors of the various HETE isomers tend to be less studied, but 12S-hydroperoxy-ETE is reported to be a key player in oxidative stress in platelets and is known to stimulate the metabolism or arachidonate and other polyunsaturated fatty acids by activating phospholipase A2 (cPLA2) and cyclooxygenase (COX-1), the first enzymes required for prostanoid production.
15S-Hydroxy-5c,8c,11c,13t-eicosatetraenoic acid (15S-HETE) is a precursor of the lipoxins and is produced by two enzymes in human tissues, one of which is related structurally to the 12-lipoxygenase of leukocytes. Indeed, this 15-lipoxygenase (15-LOX-1) is unusual in that it produces some 12-HETE in addition to the 15‑isomer. The second form of the enzyme was first found in the epidermis, although it is now know to exist in other tissues. 15S-HETE has been implicated in cell differentiation, inflammation, asthma, carcinogenesis and atherogenesis. In particular, there is an accumulation of 15-HETE in human carotid plaques, and this is believed to play a role in the induction of atherothrombotic events by increasing platelet aggregation and thrombin generation. 15-HETE appears to contribute to the development of Hodgkin lymphoma, colorectal and many other cancers, but that produced by 15‑LOX‑2 activates PPARγ, a nuclear transcription factor involved in epithelial differentiation, which may explain an anti-proliferative action on prostate cancer cells. The activity of 15-LOX-1 is important for the processes of apoptosis, autophagy and ferroptosis, and reduced levels of this enzyme in some cancers leads to decreased activity of PPARγ, resulting in loss of apoptosis and enhanced cell proliferation.
Of the terminal and near-terminal HETE isomers (cytochrome P450 metabolites), 20-HETE is pro-inflammatory and has largely detrimental functions, for example in increasing hypertension, in promoting systemic vasoconstriction and in tumour growth; it may be a factor in rheumatoid arthritis. It regulates vascular smooth muscle and endothelial cells by influencing their proliferation, migration, survival, and tube formation, acting via a specific G protein receptor (GPR75). In contrast, 20-HETE exerts a beneficial effect in terms of insulin secretion, and in the kidney, it has anti-hypertensive effects by blocking re-absorption of sodium by inhibiting the Na+‑K+‑ATPase, although it has been implicated in the pathogenesis of other kidney diseases. Production of 20-HETE is increased after the onset of both ischemic and hemorrhagic strokes. 20‑HETE may promote tumour growth, but 8- and 11‑HETE have anti-tumour activities. EPA and DHA are potent inhibitors of the biosynthesis of this compound, suggesting that this may be a partial explanation for the physiological role of omega-3 fatty acids. Other HETE isomers appear to act in opposition to 20-HETE, and 18- and 19-HETE, for example, induce vasodilatation by inhibiting the effects of 20‑HETE. In addition, they together with 16- and 17‑HETE induce re-uptake of sodium in the kidney, and 16‑HETE inhibits neutrophil adhesion so may be important in inflammation.
The 3-hydroxy-eicosanoids produced by pathogenic fungi may play a role in the inflammatory processes associated with infections by such organisms, as they are strong pro-inflammatory lipid mediators. As they are produced during the reproductive phase of yeast and fungal growth, they may also be important for the organism per se.
Metabolites of EPA and DHA: The EPA lipoxygenase metabolite 5-hydroxy-eicosapentaenoic acid (5-HEPE) enhances the induction of regulatory T cells (Tregs) that modulate the immune system and prevent autoimmune disease, and it is able to increase insulin secretion from pancreatic beta cells in mice. 12-HEPE is induced in brown fat when exposed to cold, and has been termed a 'batokine' or 'lipokine' that improves glucose metabolism by promoting glucose uptake into adipocytes and skeletal muscle through activation of an insulin-like intracellular signalling pathway. Various HEPE isomers have been detected in psoriatic arthritis, where that are believed to have anti-inflammatory effects. Apart from its function as a precursor of E-series resolvins, 18-HEPE per se has cardioprotective properties and inhibits metastasis in a cancer model.
It is noteworthy that while 17‑hydroxy-DHA derived from the action of 15-LOX is often considered simply as a precursor of the specialized pro-resolving mediators, there is evidence that it rather than the latter is important in alleviating the sensitivity to heat pain and to osteoarthritis pain in humans. In obese mice, 17‑HDHA attenuated inflammation in adipose tissue and improved insulin sensitivity and glucose tolerance.
Epoxyeicosatrienoic acids (EETs): As the regioisomers and enantiomeric forms have many similar metabolic and functional properties, epoxyeicosatrienoic acids have often been treated as a single class of compounds, although as knowledge has expanded this view is no longer justifiable. 11,12-EET in particular has a number of distinctive activities. The various EETs have major functions as autocrine and paracrine effectors in the cardiovascular and renal systems, which are believed to be largely beneficial.
Because of the anti-hypertensive, fibrinolytic, and anti-thrombotic properties of EETs, their presence in red blood cells has important implications for the control of circulation and the physical properties of the circulating blood. Both cis- and trans-EETs are synthesised and stored in erythrocytes, and they are produced and released in response to a low oxygen concentration as during exercise, for example. 11,12-EET enhances the process by which immature precursor cells develop into mature blood cells (hematopoiesis) and their further development (engraftment) in mice and zebrafish in vitro. In the kidney, EETs modulate ion transport and gene expression, producing vasodilation. They exert beneficial effects on insulin resistance and obesity-associated diseases, they alleviate inflammatory pain, neuroinflammation and neuroinflammatory diseases, they improve lung function and wound healing, and they have the potential to combat cardiovascular diseases. There is a suggestion that signalling by 11,12-EET may be a factor in the regulation of the response to DNA damage, and it is reported to beneficial towards pulmonary fibrosis. As significant amounts of EETs are incorporated into phospholipids from which they are rapidly released in the presence of Ca2+ ionophores, it has been suggested that they may be involved in those signal transduction processes mediated by phospholipases.
Some of the activities of epoxy-eicosanoids may require cell-surface receptors, and GPR132 has been identified as a potentially low-affinity EET receptor with physiological relevance in hematopoiesis, but other activities involve intracellular mechanisms, i.e. direct interaction with ion channels, signalling proteins or transcription factors. In the central nervous system, epoxyeicosanoids may have additional functions, for example in the regulation of the release of neurohormones and neuropeptides. By reducing the long-term damage associated with central neurologic insults, they may have beneficial effects towards neurologic diseases including Parkinson's disease, Alzheimer's disease and dementia. Their concentrations are controlled by soluble epoxide hydrolases, and it is hoped that inhibitors of these will be developed with therapeutic potential against a number of debilitating inflammatory diseases. In non-failing human hearts, one isoform of phospholipase A (cPLA2ζ) channels arachidonic acid into protective EETs, whereas in failing hearts, opening of the mitochondrial permeability transition pore increases the activity of a second isoform of phospholipase A (cPLA2γ) that channels arachidonic acid into toxic HETEs.
17,18-Epoxyeicosatetraenoic acid is the main epoxide regioisomer synthesised from eicosapentaenoic acid and has anti-allergy and anti-inflammatory properties by activating the receptor GPR40; it may have therapeutic properties in the skin and intestines. In sensory neurons, it functions through the prostacyclin receptor (IP) to sensitize the transient receptor potential vanilloid 1 (TRPV1). It is a vasodilator and may be responsible for some of the beneficial effects of dietary omega-3 fatty acids. Similarly, 19,20-epoxy-docosapentaenoic acid, derived from DHA, has been shown to have a number of beneficial functions in tissues. However, much less is known of the function of the other oxygenated metabolites of EPA and DHA. They appear to act in opposition to HETE isomers and may be especially important in the cardiovascular system and as anti-cancer agents.
Until recently, EETs were considered to be relatively benign molecules, but it has now been demonstrated in mice that they are stimulants for the release from dormancy of primary cancer tumours, for promoting their growth and for triggering metastasis, i.e. the spread of cancer to other organs. Their dihydroxy metabolites have even stronger effects upon cancer progression. The dihydroxyeicosatrienoic acids (DHET) produced by epoxide hydrolases are pro-inflammatory in general, and in particular they have been associated with colonic inflammation in obese mice and with osteoarthritis in the knee in humans. 19,20‑Dihydroxydocosapentaenoic acid has an important role in the development of diabetic retinopathy. As dihydroxy metabolites may also be involved in psychiatric and neurological disorders, the soluble epoxide hydrolase (EPHX1) is considered to be a therapeutic target for such conditions, as well as to assist cardiac recovery after ischemia. Inhibitors of the enzyme are undergoing clinical trials for several inflammatory conditions, including cancer.
Octadecanoids: The oxidized linoleate metabolites 13(S)-HODE and 9(S)-HODE are believed to be atherogenic through the induction of pro-inflammatory cytokines and by formation of foam cells from macrophages by activation of PPARs and other receptors. They are found at increased levels in psoriatic skin and they contribute to hepatic injury, especially non-alcoholic steatohepatitis. 9(S)-HODE is a marker for oxidative stress and contributes to the process of pain perception. In a Drosophila model, it functions by regulating FOXO family transcription factors. As the actions of octadecanoids on the regulation of inflammation are of relevance to the metabolic processes associated with atherogenesis and cancer, they are attracting special interest. They may also react non-enzymatically with proteins to form potentially toxic adducts.
In contrast, there is evidence that a 15-LOX metabolite 13S-HPODE induces apoptosis in colon cancer cells. 13(S)-HODE is the brains of rat pups is reported to increase axonal outgrowth cortical neurons in male rat pups significantly, but not in female pups where linoleic acid per se displayed this activity.
The epoxides derived from linoleic acid have been associated with multiple organ failure and in adult respiratory distress syndrome in burn patients, although it is their dihydroxy-metabolites that are the main reason for the toxicity. For example, 12,13‑dihydroxy-9Z-octadecenoic acid (12,13-diHOME) causes increased sensitivity to inflammatory pain. On the other hand, synthesis of this metabolite is induced by cold with the effect of stimulating the activity of brown adipose tissue by promoting the uptake of fatty acids. Plasma levels are increased by exercise and the source is believed to be brown adipose tissue. Therefore, 12,13-diHOME is considered to be an exercise-stimulated lipokine that produces an increase in fatty acid oxidation and uptake in skeletal muscle that results in improved whole-body metabolic homeostasis. It has been detected in human milk where it is believed to influence infant adiposity.
Although they are especially important in skin, where they are vital for epidermal barrier formation (and some argue for "mammalian survival"), the levels of hepoxilin-like triHOMEs have been found to be dysregulated in asthma and chronic obstructive pulmonary disease (COPD).
Oxidized phospholipids: Although studies are at a relatively early stage, it is becoming apparent that esterified HETEs may have specific biological functions of their own, especially in relation to apoptosis, immune regulation, signalling and blood coagulation. For example, oxidation of cardiolipin by a specific enzyme of the cytochrome c family is an important event in triggering mitochondrial apoptosis, while phosphatidylserine containing oxidized fatty acids that is externalized to the surface of cells is an especially effective signal for engulfment and digestion of apoptotic cells. Phosphatidylethanolamines containing arachidonic and adrenic acids that are oxidized by 15-LOX are believed to be pro-ferroptotic signals, but oxidized phosphatidylcholine is reported to have anti-inflammatory and protective effects in the lung. 12(S)-HETE-lysophospholipids react very specifically with certain human monocytes to generate tumour necrosis factor α (TNFα) and thence initiate a key signalling pathway; they may serve as biomarkers for age-related diseases and could potentially be used as targets for therapeutic intervention. Indeed, dys-regulation of the metabolism of any of these molecules may have implications for human health. Biological effects of phospholipids produced non-enzymatically are discussed in our web page on oxidized phospholipids.
The remarkable array of eicosanoids and related oxygenated metabolites in animal tissues provides a daunting task for analysts. Selective extraction, concentration and derivatization steps are required, followed by gas chromatography or high-performance liquid chromatography linked to mass spectrometry. Of the many published procedures, the most comprehensive appears to be that of Wang et al. cited below, who describe the analysis of 184 distinct eicosanoids in a single chromatographic run in only five minutes, with the use of deuterated internal standards and tandem mass spectrometry to ensure accurate quantification (I have no personal experience in this area).
- An, J.U., Kim, S.E. and Oh, D.K. Molecular insights into lipoxygenases for biocatalytic synthesis of diverse lipid mediators. Prog. Lipid Res., 83, 101110 (2021); DOI.
- Biringer, R.G. The enzymology of human eicosanoid pathways: the lipoxygenase branches. Mol. Biol. Rep., 47, 7189-7207 (2020); DOI.
- Biringer, R.G. A review of non-prostanoid, eicosanoid receptors: expression, characterization, regulation, and mechanism of action. J. Cell Commun. Signal., in press (2021); DOI.
- Capdevila, J.H. and Falck, J.R. The arachidonic acid monooxygenase: from biochemical curiosity to physiological/pathophysiological significance. J. Lipid Res., 59, 2047-2062 (2018); DOI.
- Evangelista, E.A., Cho, C.W., Aliwarga, T. and Totah, R.A. Expression and function of eicosanoid-producing cytochrome P450 enzymes in solid tumors. Front. Pharm., 11, 828 (2020); DOI.
- Hammerer, L., Winkler, C.K. and Kroutil, W. Regioselective biocatalytic hydroxylation of fatty acids by cytochrome P450s. Catal. Letts, 148, 787-812 (2018); DOI.
- Hajeyah, A.A., Griffiths, W.J., Wang, Y., Finch, A.J. and O’Donnell, V.B. The biosynthesis of enzymatically oxidized lipids. Front. Endocrinol., 11, 591819 (2020); DOI.
- Hildreth, K., Kodani, S.D., Hammock, B.D. and Zhao, L. Cytochrome P450-derived linoleic acid metabolites EpOMEs and DiHOMEs: a review of recent studies. J. Nutr. Biochem., 86, 108484 (2020); DOI.
- Lagarde, M. and Nicolaou, A. (Editors) Oxygenated metabolism of PUFA: analysis and biological relevance. Biochim. Biophys. Acta, Lipids (Volume 1851, Issue 4, Pages 307-518) (2015) - special issue.
- Mashima, R. and Okuyama, T. The role of lipoxygenases in pathophysiology; new insights and future perspectives. Redox Biol., 6, 297-310 (2015); DOI.
- Mendoza, S.R., Zamith-Miranda, D., Takács, T., Gacser, A., Nosanchuk, J.D. and Guimarães, A.J. Complex and controversial roles of eicosanoids in fungal pathogenesis. J. Fungi, 7, 254 (2021); DOI.
- Newcomer, M.E. and Brash, A.R. The structural basis for specificity in lipoxygenase catalysis. Protein Sci., 24, 298-309 (2015); DOI.
- Sarparast, M., Dattmore, D., Alan, J. and Lee, K.S.S. Cytochrome P450 metabolism of polyunsaturated fatty acids and neurodegeneration. Nutrients, 12, 3523 (2020); DOI.
- Shoieb, S.M., El-Sherbeni, A.A. and El-Kadi, A.O.S. Subterminal hydroxyeicosatetraenoic acids: Crucial lipid mediators in normal physiology and disease states. Chemico-Biol. Interactions, 299, 140-150 (2019); DOI.
- Singh, N.K. and Rao, G.N. Emerging role of 12/15-Lipoxygenase (ALOX15) in human pathologies. Prog. Lipid Res., 73, 28-45 (2019); DOI.
- Spector, A.A. and Kim, H.-Y. Cytochrome P450 epoxygenase pathway of polyunsaturated fatty acid metabolism. Biochim. Biophys. Acta, Lipids, 1851, 356-365 (2015); DOI.
- Tyurina, Y.Y. and 16 others. "Only a life lived for others is worth living": redox signaling by oxygenated phospholipids in cell fate decisions. Antiox. Redox Signal., 29, 1333-1358 (2018); DOI.
- Wang, B., Wu, L.J., Chen, J., Dong, L.L., Chen, C., Wen, Z., Hu, J., Fleming, I. and Wang, D.W. Metabolism pathways of arachidonic acids: mechanisms and potential therapeutic targets. Signal Transd. Targ. Ther., 6, 94 (2021); DOI.
- Wang, Y., Armando, A.M., Quehenberger, O., Yan, C. and Dennis, E.A. Comprehensive ultra-performance liquid chromatographic separation and mass spectrometric analysis of eicosanoid metabolites in human samples. J. Chromatogr. A, 1359, 60-69 (2014); DOI.
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