The crystal structure of Pseudomonas aeruginosa lipoxygenase Ala420Gly mutant explains the improved oxygen affinity and the altered reaction specificity

Kalms, Jacqueline; Banthiya, Swathi; Yoga, Etienne Galemou; Hámberg, Mats; Holzhütter, Hermann‐Georg; Kühn, Hartmut; Scheerer, Patrick

doi:10.1016/j.bbalip.2017.01.003

Cited by 29 publications

(42 citation statements)

References 72 publications

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“…The oxygen affinities of different LOX isoforms are rather high and oxygen Km values in the lower micromolar range have been reported (Egmond, Brunori, & Fasella, ; Juranek, Suzuki, & Yamamoto, ; Knapp & Klinman, ; Ludwig et al, ). On the other hand, wild‐type P. aeruginosa LOX has a Km for oxygen of about 0.4 mM indicating that under normoxic conditions this enzyme does not work at substrate saturation (Kalms et al, ). To estimate the oxygen affinity of MF‐LOX1, we performed activity assays at different oxygen concentrations and quantified the amounts of conjugated dienes formed during a 1‐min oxygenation period.…”

Section: Resultsmentioning

confidence: 99%

“…(d) Oxygen sensing: The oxygen affinity of most mammalian LOXs varies between 3 and 30 μM (Juranek et al, ). However, PA‐LOX (Kalms et al, ) exhibits a low oxygen affinity (Km > 400 μM). Since such kinetic properties are characteristic for sensor proteins (Berra et al, ), PA‐LOX might be involved in oxygen sensing.…”

Section: Discussionmentioning

confidence: 99%

“…On the other hand, wild-type P. aeruginosa LOX has a Km for oxygen of about 0.4 mM indicating that under normoxic conditions this enzyme does not work at substrate saturation (Kalms et al, 2017). To estimate the oxygen affinity of MF-LOX1, we performed activity assays at different oxygen concentrations and quantified the amounts of conjugated dienes formed during a 1-min oxygenation period.…”

Section: Enzymatic Characterization Of Mf-lox1mentioning

confidence: 99%

“…(d)Oxygen sensing: The oxygen affinity of most mammalian LOXs varies between 3 and 30 μM(Juranek et al, 1999). However, PA-LOX(Kalms et al, 2017) exhibits a low oxygen affinity (Km > 400 μM).…”

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confidence: 99%

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Functional characterization of a novel arachidonic acid 12S‐lipoxygenase in the halotolerant bacterium Myxococcus fulvus exhibiting complex social living patterns

et al. 2018

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Lipoxygenases are lipid peroxidizing enzymes, which frequently occur in higher plants and mammals. These enzymes are also expressed in lower multicellular organisms but here they are not widely distributed. In bacteria, lipoxygenases rarely occur and evaluation of the currently available bacterial genomes suggested that <0.5% of all sequenced bacterial species carry putative lipoxygenase genes. We recently rescreened the public bacterial genome databases for lipoxygenase‐like sequences and identified two novel lipoxygenase isoforms ( MF‐LOX1 and MF‐LOX2 ) in the halotolerant Myxococcus fulvus . Both enzymes share a low degree of amino acid conservation with well‐characterized eukaryotic lipoxygenase isoforms but they involve the catalytically essential iron cluster. Here, we cloned the MF‐LOX1 cDNA, expressed the corresponding enzyme as N‐terminal hexa‐his‐tag fusion protein, purified the recombinant enzyme to electrophoretic homogeneity, and characterized it with respect to its protein‐chemical and enzymatic properties. We found that M. fulvus expresses a catalytically active intracellular lipoxygenase that converts arachidonic acid and other polyunsaturated fatty acids enantioselectively to the corresponding n‐9 hydroperoxy derivatives. The enzyme prefers C 20 ‐ and C 22 ‐polyenoic fatty acids but does not exhibit significant membrane oxygenase activity. The possible biological relevance of MF‐LOX1 will be discussed in the context of the suggested concepts of other bacterial lipoxygenases.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Enzymatic Characterization Of Mf-lox1mentioning

confidence: 99%

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Functional characterization of a novel arachidonic acid 12S‐lipoxygenase in the halotolerant bacterium Myxococcus fulvus exhibiting complex social living patterns

et al. 2018

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“…The crystal structures of 15LO2 (4nre) and PEBP1 (1beh) were used to build the coarse-grained models. For LoxA, the crystal structure 5lc8 (Banthiya et al, 2016; Kalms et al, 2017) was used. Two simulations were set up, one in which the PEBP1 was place 2.5 nm above the helical bundle, of the 15LO2 (or LoxA) protein and another in which it was place below the alpha-helical bundle, away from the N-terminal beta-barrel (or N-terminal helix in LoxA).…”

Section: Star Methods Textmentioning

confidence: 99%

PEBP1 Wardens Ferroptosis by Enabling Lipoxygenase Generation of Lipid Death Signals

Wenzel

Tyurina

Zhao

et al. 2017

Cell

680

606

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SUMMARY Ferroptosis is a form of programmed cell death pathogenic to several acute and chronic diseases and executed via oxygenation of polyunsaturated phosphatidylethanolamines (PE) by 15-lipoxygenases (15-LO) that normally use free polyunsaturated fatty acids as substrates. Mechanisms of the altered 15-LO substrate specificity are enigmatic. We sought a common ferroptosis regulator for 15LO. We discovered that PEBP1, a scaffold protein inhibitor of protein kinase cascades, complexes with two 15LO isoforms, 15LO1 and 15LO2, and changes their substrate competence to generate hydroperoxy-PE. Inadequate reduction of hydroperoxy-PE due to insufficiency or dysfunction of a selenoperoxidase, GPX4, leads to ferroptosis. We demonstrated the importance of PEBP1-dependent regulatory mechanisms of ferroptotic death in airway epithelial cells in asthma, kidney epithelial cells in renal failure and cortical and hippocampal neurons in brain trauma. As master regulators of ferroptotic cell death with profound implications for human disease, PEBP1/15LO complexes represent a new target for drug discovery.

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Thirty years with three-dimensional structures of lipoxygenases

Oliw

2024

Archives of Biochemistry and Biophysics

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The crystal structure of Pseudomonas aeruginosa lipoxygenase Ala420Gly mutant explains the improved oxygen affinity and the altered reaction specificity

Cited by 29 publications

References 72 publications

Functional characterization of a novel arachidonic acid 12S‐lipoxygenase in the halotolerant bacterium Myxococcus fulvus exhibiting complex social living patterns

Functional characterization of a novel arachidonic acid 12S‐lipoxygenase in the halotolerant bacterium Myxococcus fulvus exhibiting complex social living patterns

PEBP1 Wardens Ferroptosis by Enabling Lipoxygenase Generation of Lipid Death Signals

Thirty years with three-dimensional structures of lipoxygenases

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