Secreted lipoxygenase from Pseudomonas aeruginosa exhibits biomembrane oxygenase activity and induces hemolysis in human red blood cells

Banthiya, Swathi; Pekárová, Mária; Kühn, Hartmut; Heydeck, Dagmar

doi:10.1016/j.abb.2015.09.003

Cited by 25 publications

(38 citation statements)

References 37 publications

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“…by guest, on May 11, 2018 www.jlr.org Cloning and expression and purifi cation of recombinant AmLOX1 in E. coli Acaryochloris marina (MBIC11017) was obtained from the NITE Biological Resource Center, Department of Biotechnology, Japan. Genomic DNA was isolated from approximately 50 mg of frozen cell material, as previously described ( 17 ).…”

Section: Methodsmentioning

confidence: 99%

“…by guest, on May 11, 2018 www.jlr.org in O ( 35 ). The structure was initially refi ned using CNS ( 36 ) against the MLHL target (maximum likelihood, HendricksonLattman coeffi cients) and verifi ed against simulated annealing omit maps, as well as SIGMA A -weighted electron density maps.…”

Section: Cyanobacterial Materials and Growth Conditionsmentioning

confidence: 99%

“…by guest, on May 11, 2018 www.jlr.org energetically favored is, in general, represented in the crystal structure and stabilized by crystal contacts. The position of the two N-terminal amphipathic ␣ -helices is further driven by contact with the catalytic domain.…”

Section: The ␣ -Helical Extension At the N-terminus Facilitates Bindimentioning

confidence: 99%

“…by guest, on May 11, 2018 www.jlr.org Replacement method was not possible. The phase problem was fi nally solved by single isomorphous replacement with anomalous scattering, using a gold derivative of CspLOX1 (supplementary Table 1).…”

Section: Structure Determination and Refi Nementmentioning

confidence: 99%

“…Finally SHARP/autoSHARP ( 33 ) was tried using the best heavy atom sites from previous SHELD runs. This resulted in obtaining an interpretable electron density map, which was used by ARP/wARP ( 34 ) to build an initial model, which was then rebuilt by guest, on May 11, 2018 www.jlr.org number WP_006102195.1) with 61% sequence identity and to a predicted LOX from the cyanobacterium Microcystis aeruginosa (GenBank accession number WP_046663104.1) with 40% sequence identity. CspLOX1, which has a calculated molecular mass of 76.5 kDa, is rather small compared with other eukaryotic LOXs, ranging from approximately 75-80 kDa for animal LOXs to 94-104 kDa for plant LOXs ( 14 ).…”

Section: Structure Determination and Refi Nementmentioning

confidence: 99%

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Crystal structure of a lipoxygenase from Cyanothece sp. may reveal novel features for substrate acquisition

Newie¹,

Andreou²,

Neumann

et al. 2016

Journal of Lipid Research

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This article is available online at http://www.jlr.org PUFA oxygenation is a process that leads to the formation of bioactive lipid compounds with a diversity of biological functions in plants and animals ( 1, 2 ). Oxidation of PUFAs may be catalyzed by two major classes of enzymes, cyclooxygenases or ␣ -dioxygenases and lipoxygenases (LOXs) ( 3 ). Of the two, LOXs are nonheme iron-containing dioxygenases that are widely found in higher plants and animals, but have also been detected in some corals, mosses, fungi, and a number of bacteria ( 4, 5 ). Members of the LOX family catalyze the regio-and stereospecifi c oxygenation of PUFAs with one or more (1 Z ,4 Z )-pentadiene moieties leading to the formation of hydroperoxy PUFAs ( 6 ). LOX hydroperoxide products are precursors of important signaling compounds such as aldehydes and jasmonates in plants and leukotrienes, resolvins, and lipoxins in mammals ( 3 ). These signaling molecules play an important role in wound and defense responses as well as in aspects of plant development ( 7 ), while in mammals they function in infl ammation, asthma, and the development of atherosclerosis and cancer ( 1 ). Other than in higher organisms, very little is still known regarding the overall function of LOX products in prokaryotes and fungi ( 4, 5 ).As the regio-and stereospecifi city of the LOX reaction has an infl uence on the biological function of the product, many studies have focused on the molecular basis of this specifi city. In the case of arachidonic acid [20:4(n-6)], which is a typical mammalian LOX substrate, several Abstract In eukaryotes, oxidized PUFAs, so-called oxylipins, are vital signaling molecules. The fi rst step in their biosynthesis may be catalyzed by a lipoxygenase (LOX), which forms hydroperoxides by introducing dioxygen into PUFAs. Here we characterized CspLOX1, a phylogenetically distant LOX family member from Cyanothece sp. PCC 8801 and determined its crystal structure. In addition to the classical two domains found in plant, animal, and coral LOXs, we identifi ed an N-terminal helical extension, reminiscent of the long ␣ -helical insertion in Pseudomonas aeruginosa LOX. In liposome fl otation studies, this helical extension, rather than the ␤ -barrel domain, was crucial for a membrane binding function. Additionally, CspLOX1 could oxygenate 1,2-diarachidonyl-sn -glycero-3-phosphocholine, suggesting that the enzyme may act directly on membranes and that fatty acids bind to the active site in a tail-fi rst orientation. This binding mode is further supported by the fact that CspLOX1 catalyzed oxygenation at the n-10 position of both linoleic and arachidonic acid, resulting in 9 R -and 11 R -hydroperoxides, respectively. Together these results reveal unifying structural features of LOXs and their function. While the core of the active site is important for lipoxygenation and thus highly conserved, peripheral domains functioning in membrane and substrate binding are more variable.

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Section: Methodsmentioning

confidence: 99%

Section: Cyanobacterial Materials and Growth Conditionsmentioning

confidence: 99%

Section: The ␣ -Helical Extension At the N-terminus Facilitates Bindimentioning

confidence: 99%

Section: Structure Determination and Refi Nementmentioning

confidence: 99%

Section: Structure Determination and Refi Nementmentioning

confidence: 99%

See 3 more Smart Citations

Crystal structure of a lipoxygenase from Cyanothece sp. may reveal novel features for substrate acquisition

Newie¹,

Andreou²,

Neumann

et al. 2016

Journal of Lipid Research

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Secretion of Pseudomonas aeruginosa Lipoxygenase by Pichia pastoris upon Glycerol Feed

Hashem

Stolterfoht

Rinnofner

et al. 2020

Biotechnology Journal

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Pseudomonas aeruginosa lipoxygenase (PaLOX) catalyzes the peroxidation of unsaturated fatty acids. Not only linoleic acid, but also linolenic acid and oleic acid are oxidized. The natural host secretes PaLOX into the periplasmic space. Herein, the aim is to secrete PaLOX to the culture supernatant of Pichia pastoris. Since protein background in the culture supernatant is typically rather low, this strategy allows for almost pure production of PaLOX applicable for the valorization of renewable fatty acids, for example for the production of green leaf volatiles. Using the CAT1 promoter system and the well‐established α‐factor signal sequence for secretion, methanol‐ and glycerol‐induced secretion are compared and the latter shows four times more LOX activity in the culture supernatant under methanol‐free conditions. In addition, secreted PaLOX is purified and the specific activity with enzyme in culture supernatant is compared. Notably, the predominant specific activity is achieved for enzyme in culture supernatant ‐ 11.6 U mg−1 ‐ reaching five times higher specific activity than purified PaLOX.

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Catalytic Multiplicity of 15-Lipoxygenase-1 Orthologs (ALOX15) of Different Species

Kühn

Karst

Heydeck

2016

Lipoxygenases in Inflammation

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Lipoxygenases (LOX) form a family of lipid peroxidizing enzymes, which have been implicated in a number of physiological processes and in the pathogenesis of inflammatory, hyperproliferative and neurodegenerative diseases. They occur in bacteria and eucarya and the human genome involves six functional LOX genes, which encode for six different LOX isoforms. One of these isoforms is ALOX15, which has first been described in rabbits in 1974 as an enzyme capable of oxidizing membrane phospholipids during the maturational breakdown of mitochondria in immature red blood cells. During the following decades ALOX15 orthologs have extensively been characterized and their biological functions have been studied in a number of cellular in vitro systems as well as in various whole animal disease models. This review is aimed at summarizing the current knowledge on the protein-chemical, molecular biological and enzymatic properties of ALOX15 orthologs of various mammalian species (rabbit, pig, human, nonhuman primates, mouse, rat). Because of space limitations the biological roles of ALOX15 orthologs have not been addressed since this topic has extensively been covered in a previous review (Kuhn et al., Biochim Biophys Acta 1851:308-330, 2015.

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Secreted lipoxygenase from Pseudomonas aeruginosa exhibits biomembrane oxygenase activity and induces hemolysis in human red blood cells

Cited by 25 publications

References 37 publications

Crystal structure of a lipoxygenase from Cyanothece sp. may reveal novel features for substrate acquisition

Crystal structure of a lipoxygenase from Cyanothece sp. may reveal novel features for substrate acquisition

Secretion of Pseudomonas aeruginosa Lipoxygenase by Pichia pastoris upon Glycerol Feed

Catalytic Multiplicity of 15-Lipoxygenase-1 Orthologs (ALOX15) of Different Species

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