Monolysocardiolipin: improved preparation with high yield

Kim, Junhwan; Hoppel, Charles L.

doi:10.1194/jlr.d010587

Cited by 11 publications

(14 citation statements)

References 23 publications

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“…B, relative amounts of 9-HODE, 13-HODE, and LA in TLCL/oxTLCL/PAPC vesicles after complete hydrolysis by porcine pancreas PLA 2 and T. lanuginosus PLA 1 , derivatization with AMPP, and analysis by LC-MS/MS. C-E, oxidized cardiolipin used as substrate for iPLA 2 ␥ hydrolysis was prepared by cytochrome c-mediated oxidation in the presence of hydrogen peroxide as described previously (52) and purified by reversed-phase HPLC (C). Fraction 1 (D) and fraction 2 (E) containing oxTLCL were collected and analyzed by mass spectrometry as described under "Experimental procedures."…”

Section: Discussionmentioning

confidence: 99%

The phospholipase iPLA2γ is a major mediator releasing oxidized aliphatic chains from cardiolipin, integrating mitochondrial bioenergetics and signaling

Liu

Moon

Jenkins

et al. 2017

Journal of Biological Chemistry

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Cardiolipin (CL) is a dimeric phospholipid with critical roles in mitochondrial bioenergetics and signaling. Recently, inhibition of the release of oxidized fatty acyl chains from CL by the calcium-independent phospholipase Aγ (iPLAγ)-selective inhibitor (R)-BEL suggested that iPLAγ is responsible for the hydrolysis of oxidized CL and subsequent signaling mediated by the released oxidized fatty acids. However, chemical inhibition by BEL is subject to off-target pharmacologic effects. Accordingly, to unambiguously determine the role of iPLAγ in the hydrolysis of oxidized CL, we compared alterations in oxidized CLs and the release of oxidized aliphatic chains from CL in experiments with purified recombinant iPLAγ, germ-line iPLAγ mice, cardiac myocyte-specific iPLAγ transgenic mice, and wild-type mice. Using charge-switch high mass accuracy LC-MS/MS with selected reaction monitoring and product ion accurate masses, we demonstrated that iPLAγ is the major enzyme responsible for the release of oxidized aliphatic chains from CL. Our results also indicated that iPLAγ selectively hydrolyzes 9-hydroxy-octadecenoic acid in comparison to 13-hydroxy-octadecenoic acid from oxidized CLs. Moreover, oxidative stress (ADP, NADPH, and Fe) resulted in the robust production of oxidized CLs in intact mitochondria from iPLAγ mice. In sharp contrast, oxidized CLs were readily hydrolyzed in mitochondria from wild-type mice during oxidative stress. Finally, we demonstrated that CL activates the iPLAγ-mediated hydrolysis of arachidonic acid from phosphatidylcholine, thereby integrating the production of lipid messengers from different lipid classes in mitochondria. Collectively, these results demonstrate the integrated roles of CL and iPLAγ in lipid second-messenger production and mitochondrial bioenergetics during oxidative stress.

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Section: Discussionmentioning

confidence: 99%

The phospholipase iPLA2γ is a major mediator releasing oxidized aliphatic chains from cardiolipin, integrating mitochondrial bioenergetics and signaling

Liu

Moon

Jenkins

et al. 2017

Journal of Biological Chemistry

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“…Standards for phospholipids and oxygenated fatty acids were from Avanti Polar Lipids and Cayman Chemicals. Standards of mCLs were prepared as described previously 43 .…”

Section: Methodsmentioning

confidence: 99%

Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice

et al. 2014

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Ferroptosis is a non-apoptotic form of cell death induced by small molecules in specific tumour types, and in engineered cells overexpressing oncogenic RAS. Yet, its relevance in non-transformed cells and tissues is unexplored and remains enigmatic. Here, we provide direct genetic evidence that the knockout of glutathione peroxidase 4 (Gpx4) causes cell death in a pathologically relevant form of ferroptosis. Using inducible Gpx4−/− mice, we elucidate an essential role for the glutathione/Gpx4 axis in preventing lipid-oxidation-induced acute renal failure and associated death. We furthermore systematically evaluated a library of small molecules for possible ferroptosis inhibitors, leading to the discovery of a potent spiroquinoxalinamine derivative called Liproxstatin-1, which is able to suppress ferroptosis in cells, in Gpx4−/− mice, and in a pre-clinical model of ischaemia/reperfusion-induced hepatic damage. In sum, we demonstrate that ferroptosis is a pervasive and dynamic form of cell death, which, when impeded, promises substantial cytoprotection.

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“…Both internal and reference standards of MLCLs were prepared from CL-(14:0/14:0/14:0/14:0) and CL-(18:2/18:2/18:2/18:2), respectively, as described. 26 Precursor ions [M–H] were identified with a mass accuracy less than 5 ppm. Therefore, phospholipids, including CL, were identified by accurate mass and MS 2 fragmentation analysis.…”

Section: Methodsmentioning

confidence: 99%

“…Liposomes containing DOPC and CL (10 μ M, 1:1) were prepared by sonication (Ultrasonic Homogenizer 4710 series, Cole-Parmer Instrument Co., Chicago, IL) and treated with Cld1 (0.1–0.5 μ g of protein per 250 μ L sample) in 50 mM HEPES at pH 7.45, containing 100 μ M DTPA for 20 min at 37 °C. After incubation with Cld1, lipids were extracted by Folch procedure and analysis of MLCL was performed using LC-ESI-MS. Internal standards of MLCLs were prepared from respective CLs in a PLA 2 (porcine pancreatic, Sigma) driven reaction as described by Kim and Hoppel 26 with light modifications.…”

Section: Methodsmentioning

confidence: 99%

Lipidomics Characterization of Biosynthetic and Remodeling Pathways of Cardiolipins in Genetically and Nutritionally Manipulated Yeast Cells

Tyurina¹,

Lou

Qu³

et al. 2016

ACS Chem. Biol.

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Cardioipins (CLs) are unique tetra-acylated phospholipids of mitochondria and define the bioenergetics and regulatory functions of these organelles. An unresolved paradox is the high uniformity of CL molecular species (tetra-linoleoyl-CL) in the heart, liver, and skeletal muscles—in contrast to their high diversification in the brain. Here, we combined liquid chromatography–mass-spectrometry-based phospholipidomics with genetic and nutritional manipulations to explore CLs’ biosynthetic vs postsynthetic remodeling processes in S. cerevisiae yeast cells. By applying the differential phospholipidomics analysis, we evaluated the contribution of Cld1 (CL-specific phospholipase A) and Taz1 (acyl-transferase) as the major regulatory mechanisms of the remodeling process. We further established that nutritional “pressure” by high levels of free fatty acids triggered a massive synthesis of homoacylated molecular species in all classes of phospholipids, resulting in the preponderance of the respective homoacylated CLs. We found that changes in molecular speciation of CLs induced by exogenous C18-fatty acids (C18:1 and C18:2) in wild-type (wt) cells did not occur in any of the remodeling mutant cells, including cld1Δ, taz1Δ, and cld1Δtaz1Δ. Interestingly, molecular speciation of CLs in wt and double mutant cells cld1Δtaz1Δ was markedly different. Given that the bioenergetics functions are preserved in the double mutant, this suggests that the accumulated MLCL—rather than the changed CL speciation—are the likely major contributors to the mitochondrial dysfunction in taz1Δ mutant cells (also characteristic of Barth syndrome). Biochemical studies of Cld1 specificity and computer modeling confirmed the hydrolytic selectivity of the enzyme toward C16-CL substrates and the preservation of C18:1-containing CL species.

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Monolysocardiolipin: improved preparation with high yield

Cited by 11 publications

References 23 publications

The phospholipase iPLA2γ is a major mediator releasing oxidized aliphatic chains from cardiolipin, integrating mitochondrial bioenergetics and signaling

The phospholipase iPLA2γ is a major mediator releasing oxidized aliphatic chains from cardiolipin, integrating mitochondrial bioenergetics and signaling

Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice

Lipidomics Characterization of Biosynthetic and Remodeling Pathways of Cardiolipins in Genetically and Nutritionally Manipulated Yeast Cells

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