Cloning and Characterization of a Bifunctional Leukotriene A4 Hydrolase from Saccharomyces cerevisiae

Kull, Filippa; Ohlson, Eva; Haeggström, Jesper Z.

doi:10.1074/jbc.274.49.34683

Cited by 28 publications

(34 citation statements)

References 31 publications

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“…Apparently, from a noncatalytic function in certain early aminopeptidases, evolution has gradually brought Asp-375 into a key role for the epoxide hydrolase reaction in LTA4H. In this context, it is interesting to note that Asp-375 is indeed present in Saccharomyces cerevisiae LTA4H, a product of an ancestral gene for mammalian LTA4H that exhibits both a leucyl aminopeptidase and a primitive epoxide hydrolase activity against LTA 4 (31,32).…”

Section: Asp-375 Is a Catalytic Residue Specifically Required For Thementioning

confidence: 99%

Leukotriene A ₄ hydrolase: Selective abrogation of leukotriene B ₄ formation by mutation of aspartic acid 375

Rudberg

Tholander

Thunnissen

et al. 2002

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

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Leukotriene A4 (LTA4, 5S-trans-5,6-oxido-7,9-trans-11,14-cis-eicosatetraenoic acid) hydrolase (LTA4H)͞aminopeptidase is a bifunctional zinc metalloenzyme that catalyzes the final and rate-limiting step in the biosynthesis of leukotriene B4 (LTB4, 5S,12R-dihydroxy-6,14-cis-8,10-trans-eicosatetraenoic acid), a classical chemoattractant and immune modulating lipid mediator. Two chemical features are key to the bioactivity of LTB4, namely, the chirality of the 12R-hydroxyl group and the cis-trans-trans geometry of the conjugated triene structure. From the crystal structure of LTA4H, a hydrophilic patch composed of Gln-134, Tyr-267, and Asp-375 was identified in a narrow and otherwise hydrophobic pocket, believed to bind LTA4. In addition, Asp-375 belongs to peptide K21, a previously characterized 21-residue active site-peptide to which LTA4 binds during suicide inactivation. In the present report we used site-directed mutagenesis and x-ray crystallography to show that Asp-375, but none of the other candidate residues, is specifically required for the epoxide hydrolase activity of LTA4H. Thus, mutation of Asp-375 leads to a selective loss of the enzyme's ability to generate LTB4 whereas the aminopeptidase activity is preserved. We propose that Asp-375, possibly assisted by Gln-134, acts as a critical determinant for the stereoselective introduction of the 12R-hydroxyl group and thus the biological activity of LTB4.

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Section: Asp-375 Is a Catalytic Residue Specifically Required For Thementioning

confidence: 99%

Leukotriene A ₄ hydrolase: Selective abrogation of leukotriene B ₄ formation by mutation of aspartic acid 375

Rudberg

Tholander

Thunnissen

et al. 2002

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

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“…For instance, aminopeptidase 1 from Caenorhabditis elegans, which is 45% identical (63% similar) at the amino acid level to mammalian LTA4H, fails to hydrolyze LTA 4 into LTB 4 (42). On the other hand, an LTA4H that is 39% identical (53% similar) to the human enzyme has been cloned and characterized from yeast, Saccharomyces cerevisiae (43). The S. cerevisiae LTA4H is a zinc leucyl aminopeptidase with a primitive epoxide hydrolase activity against LTA 4 .…”

Section: Molecular Evolution Of Lta 4 Hydrolasementioning

confidence: 99%

Leukotriene A4 Hydrolase/Aminopeptidase, the Gatekeeper of Chemotactic Leukotriene B4 Biosynthesis

Haeggström

2004

Journal of Biological Chemistry

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The leukotrienes (LTs) 1 are a family of lipid mediators that play important roles in a variety of allergic and inflammatory reactions (1, 2). These molecules are formed by leukocytes and are divided into two classes, the spasmogenic cysteinyl leukotrienes and LTB 4 , which is a classical chemoattractant that triggers adherence and aggregation of leukocytes to the endothelium at nanomolar concentrations. Recent data also indicate that LTB 4 is a chemoattractant for T-cells, creating a functional link between early innate and late adaptive immune responses to inflammation (3-5). In addition, LTB 4 participates in the host defense against infections (6) and is a key mediator of platelet-activating factor-induced lethal shock (7). Because of these powerful biological effects, LTB 4 is regarded as an important chemical mediator in a variety of acute and chronic inflammatory diseases, e.g. nephritis, arthritis, dermatitis, and chronic obstructive pulmonary disease (8). Moreover, only recently, several lines of pharmacological, morphological, biochemical, and genetic evidence have been gathered implicating LTs, in particular LTB 4 , as a mediator of vascular inflammation and arteriosclerosis (9). This article gives an overview of the biochemical, structural, and catalytic properties of LTA 4 hydrolase (LTA4H), which catalyzes the final and committed step in LTB 4 biosynthesis. LTA 4 Hydrolase Is a Key Enzyme in the 5-Lipoxygenase PathwayIn cellular biosynthesis of LTs, 5-lipoxygenase, assisted by 5-lipoxygenase-activating protein, converts arachidonic acid into the unstable epoxide LTA 4 , which in turn may be enzymatically conjugated with GSH to form LTC 4 , the parent compound of the cysteinyl leukotrienes, or hydrolyzed into LTB 4 by LTA4H (Fig. 1). Leukotrienes can also be formed via transcellular routes, where LTA 4 is donated from an activated leukocyte to a recipient cell for further metabolism by downstream enzymes, a process that was recently shown to occur in vivo (10). LTB 4 signals via a specific, high affinity, G-protein-coupled receptor (BLT 1 ) (11). In addition, a second receptor for LTB 4 (BLT 2 ) has been discovered, the functional role of which is presently not known (12). Interestingly, LTB 4 is also a natural ligand of the peroxisome proliferator-activated receptor ␣ class of nuclear receptors and has been suggested to play a role in lipid homeostasis (13). Leukotriene A 4 Hydrolase Is a Zinc-dependent Epoxide Hydrolase and AminopeptidaseLTA4H is a monomeric soluble protein that is widely distributed and has been purified from several mammalian sources (14). It resides in the cytosol, although nuclear localization has also been reported recently (15). The cDNAs encoding the human, mouse, rat, and guinea pig enzymes have been cloned and sequenced. The proteins contain 610 amino acids, excluding the first Met, and the calculated molecular mass of the human enzyme is 69,153 Da. The primary, secondary, and tertiary structures of LTA4H bear no resemblance to soluble xenobiotic epoxide hydrolase, alth...

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“…A lipoxygenase was purified from Saccharomyces cerevisiae but was never cloned or sequenced (122). However, a bifunctional LTA 4 hydrolase from Saccharomyces cerevisiae has been cloned and characterized (72). This enzyme has 42% identity with mammalian LTA 4 hydrolase and produces LTB 4 from LTA 4 in vitro (73).…”

Section: Protozoa Helminths and Fungimentioning

confidence: 99%

Production of Eicosanoids and Other Oxylipins by Pathogenic Eukaryotic Microbes

2003

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Oxylipins are oxygenated metabolites of fatty acids. Eicosanoids are a subset of oxylipins and include the prostaglandins and leukotrienes, which are potent regulators of host immune responses. Host cells are one source of eicosanoids and oxylipins during infection; however, another potential source of eicosanoids is the pathogen itself. A broad range of pathogenic fungi, protozoa, and helminths produce eicosanoids and other oxylipins by novel synthesis pathways. Why do these organisms produce oxylipins? Accumulating data suggest that phase change and differentiation in these organisms are controlled by oxylipins, including prostaglandins and lipoxygenase products. The precise role of pathogen-derived eicosanoids in pathogenesis remains to be determined, but the potential link between pathogen eicosanoids and the development of TH2 responses in the host is intriguing. Mammalian prostaglandins and leukotrienes have been studied extensively, and these molecules can modulate Th1 versus Th2 immune responses, chemokine production, phagocytosis, lymphocyte proliferation, and leukocyte chemotaxis. Thus, eicosanoids and oxylipins (host or microbe) may be mediators of a direct host-pathogen “cross-talk” that promotes chronic infection and hypersensitivity disease, common features of infection by eukaryotic pathogens

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Cloning and Characterization of a Bifunctional Leukotriene A4 Hydrolase from Saccharomyces cerevisiae

Cited by 28 publications

References 31 publications

Leukotriene A ₄ hydrolase: Selective abrogation of leukotriene B ₄ formation by mutation of aspartic acid 375

Leukotriene A ₄ hydrolase: Selective abrogation of leukotriene B ₄ formation by mutation of aspartic acid 375

Leukotriene A4 Hydrolase/Aminopeptidase, the Gatekeeper of Chemotactic Leukotriene B4 Biosynthesis

Production of Eicosanoids and Other Oxylipins by Pathogenic Eukaryotic Microbes

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Cloning and Characterization of a Bifunctional Leukotriene A4 Hydrolase from Saccharomyces cerevisiae

Cited by 28 publications

References 31 publications

Leukotriene A 4 hydrolase: Selective abrogation of leukotriene B 4 formation by mutation of aspartic acid 375

Leukotriene A 4 hydrolase: Selective abrogation of leukotriene B 4 formation by mutation of aspartic acid 375

Leukotriene A4 Hydrolase/Aminopeptidase, the Gatekeeper of Chemotactic Leukotriene B4 Biosynthesis

Production of Eicosanoids and Other Oxylipins by Pathogenic Eukaryotic Microbes

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Leukotriene A ₄ hydrolase: Selective abrogation of leukotriene B ₄ formation by mutation of aspartic acid 375

Leukotriene A ₄ hydrolase: Selective abrogation of leukotriene B ₄ formation by mutation of aspartic acid 375