Investigation of calcium-dependent activity and conformational dynamics of zebra fish 12-lipoxygenase

Mittal, Monica; Hasan, Mahmudul; Balagunaseelan, Navisraj; Fauland, Alexander; Wheelock, Craig E.; Rådmark, Olof; Haeggström, Jesper Z.; Rinaldo-Matthis, Agnes

doi:10.1016/j.bbagen.2017.05.015

Cited by 6 publications

(6 citation statements)

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“…Small angle X-ray scattering indicates a high degree of flexibility between the PLAT and catalytic domains. , However, the crystal structure and scattering data reveal little about the role that structural dynamics play in the translocation of 15-LOX-2 to the membrane, regulatory mechanisms, or allosteric interactions. In this report, we investigated the structural changes in 15-LOX-2 upon binding of Ca 2+ and ligands, as well as upon membrane association using hydrogen–deuterium exchange mass spectrometry (HDX-MS).…”

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confidence: 99%

Structural Dynamics of 15-Lipoxygenase-2 via Hydrogen–Deuterium Exchange

et al. 2017

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Eicosanoids are inflammatory signaling lipids that are biosynthesized in response to cellular injury or threat. They were originally thought to be pro-inflammatory molecules, but members of at least one subclass, the lipoxins, are able to resolve inflammation. One step in lipoxin synthesis is the oxygenation of arachidonic acid by 15-lipoxygenase (15-LOX). 15-LOX contains two domains: a Ca2+ binding PLAT domain and a catalytic domain. 15-LOX is a soluble cytosolic protein until binding of Ca2+ to the PLAT domain promotes translocation to the membrane surface. The role of 15-LOX structural dynamics in this translocation has remained unclear. We investigated the dynamics of 15-LOX isoform B (15-LOX-2) upon binding of Ca2+ and ligands, as well as upon membrane association using hydrogen–deuterium exchange mass spectrometry (HDX-MS). We used HDX-MS to probe the solvent accessibility and backbone flexibility of 15-LOX-2, revealing significant differences in deuterium incorporation between the PLAT and catalytic domains, with the PLAT domain demonstrating higher flexibility. Comparison of HDX for 15-LOX-2 in the presence and absence of Ca2+ indicates there are few differences in structural dynamics. Furthermore, our HDX results involving nanodisc-associated 15-LOX-2 suggest that significant structural and dynamic changes in 15-LOX-2 are not required for membrane association. Our results also show that a substrate lipid binding to the active site in the catalytic domain does induce changes in incorporation of deuterium into the PLAT domain. Overall, our results challenge the previous hypothesis that Ca2+ binding induces major structural changes in the PLAT domain and support the hypothesis that is interdomain communication in 15-LOX-2.

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confidence: 99%

Structural Dynamics of 15-Lipoxygenase-2 via Hydrogen–Deuterium Exchange

et al. 2017

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“…118 LTs and LXs are generated by the LOX pathway, which plays a significant part in the occurrence of inflammation, 119 including many main molecules such as LOX-5, LOX-8, LOX-12 and LOX-15 in mammals, and the three LOX subtypes in fish, that is, LOX-1 (a homologue of the human LOX-15), LOX-2 (a homologue of the human LOX-5) and LOX-12. 9 Tian et al demonstrated that the LOX pathway might be the key regulator when AA is accelerated in the early stage of grass carp adipocytes. 120 It is worth noting that antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and ascorbic acid can inhibit the LOX activity in muscle of mackerel (Scomber scombrus).…”

Section: Fish Inflammationmentioning

confidence: 99%

“…AA can generate prostaglandins (PGs), leukotrienes (LTs), lipoxins (LXs) and other metabolites and regulate inflammatory response through the cyclooxygenase (COX) and lipoxygenase (LOX) pathways. [6][7][8][9][10] It has been suggested that AA plays an important role in fish inflammation.…”

Section: Introductionmentioning

confidence: 99%

“…In the presence of cellular damage, phospholipase A 2 (PLA 2 ) and phospholipase C (PLC) are activated to prompt the release of arachidonic acid (AA) from plasma membrane phospholipids. AA can generate prostaglandins (PGs), leukotrienes (LTs), lipoxins (LXs) and other metabolites and regulate inflammatory response through the cyclooxygenase (COX) and lipoxygenase (LOX) pathways 6–10 . It has been suggested that AA plays an important role in fish inflammation.…”

Section: Introductionmentioning

confidence: 99%

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The role of ferroptosis in fish inflammation

Gao

Shi

Pei

et al. 2022

Reviews in Aquaculture

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Ferroptosis, as one of cell death modalities, is driven by iron‐dependent phospholipid peroxidation and regulated by various cellular metabolic pathways, including iron, amino acid and lipid metabolism, as well as disease‐related signalling pathways. Cell survival or death is a critical issue in the inflammatory response. On the one hand, ferroptosis is more immunogenic than apoptosis, and ruptured ferroptotic cells release damage‐associated molecular patterns (DAMPs), which can trigger an inflammatory response. On the other hand, various pathogenic bacteria may cause peroxidation of polyunsaturated fatty acids (PUFAs) on the cell membrane, leading to ferroptosis. In fish, the key genes and signalling pathways related to ferroptosis are conserved, but there are still differences among fish species. In recent years, frequent occurrences of fish diseases have posed a considerable challenge to the large‐scale aquaculture. The specific precursors for ferroptotic cell death were iron‐dependent oxidative stress and lipid peroxidation, which always occur in fish inflammation. This review summarizes recent studies on ferroptosis in fish inflammation, that is, iron‐dependent programmed cell death, to address ferroptosis, inflammation and correlations between them in fish. This review may contribute to a better understanding of the underlying mechanisms of ferroptosis and inflammation in fish and provides suggestions for sustainable development and large‐scale aquaculture farming.

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“…The latter structure revealed the arachidonic acid substrate positioned in the U-shaped active site in a pose consistent with the regio- and stereo-specificity of 8 R- LOX, and hence provided a robust model for understanding LOX product specificity in this super-family (Newcomer & Brash, 2015). The protein engineering strategy of removing a LOX membrane-insertion loop has been applied to additional lipoxygenases (Mittal et al, 2017) and yielded a protein amenable to crystallization (Gilbert et al, 2011). …”

Section: Introductionmentioning

confidence: 99%

Expression of an 8 R -Lipoxygenase From the Coral Plexaura homomalla

Gilbert

Neau

Newcomer

2018

Marine Enzymes and Specialized Metabolism - Part B

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Methods are presented for the use of the coral 8R-lipoxygenase from the Caribbean sea whip coral Plexaura homomalla as a model enzyme for structural studies of animal lipoxygenases. The 8R-lipoxygenase is remarkably stable and can be stored at 4°C for 3 months with virtually no loss of activity. In addition, an engineered "pseudo wild-type" enzyme is soluble in the absence of detergents, which helps facilitate the preparation of enzyme:substrate complexes.

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Investigation of calcium-dependent activity and conformational dynamics of zebra fish 12-lipoxygenase

Cited by 6 publications

References 57 publications

Structural Dynamics of 15-Lipoxygenase-2 via Hydrogen–Deuterium Exchange

Structural Dynamics of 15-Lipoxygenase-2 via Hydrogen–Deuterium Exchange

The role of ferroptosis in fish inflammation

Expression of an 8 R -Lipoxygenase From the Coral Plexaura homomalla

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