Profiling of sphingolipids in Caenorhabditis elegans by two-dimensional multiple heart-cut liquid chromatography – mass spectrometry

Scholz, Johannes; Helmer, Patrick O.; Nicolai, Merle M.; Bornhorst, Julia; Hayen, Heiko

doi:10.1016/j.chroma.2021.462481

Cited by 11 publications

(14 citation statements)

References 45 publications

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“…Using a combination of reverse genetics, comparative lipidomics, ceramide profiling by ESI-(+)-HR-MS e , and isotope incorporation experiments with bacterial mutants specifically enriched with stable isotope labeled branched-chain amino acids (BCAA) and branchedchain fatty acids (BCFA) we characterize the biosynthetic connections between iso-fatty acid (13 -16) metabolism and ceramide (18) biosynthesis. Our results demonstrate that ceramide (18) biosynthesis proceeds from L-leucine (10) through elo-5 dependent de novo lipogenesis of C17iso-CoA (15a) and chain shortening to C15iso-CoA (14a) during the peroxisomal β-oxidation cycle catalyzed by the 3-ketoacyl-CoA thiolase daf-22, placing β-oxidation upstream of ceramide biosynthesis.…”

Section: Introductionmentioning

confidence: 75%

“…Ceramide (18) profiling of the Caenorhabditis elegans lipidome was performed by targeted HPLC-ESI-(+)-HR-MS analysis for previously described components (Figure 1,a and 1,b) [16,19] all of which were also detected upon ESI-(+)-HR-MS e screening (Figure 1,c) for the characteristic sphingosine-derived fragment ion at m/z 250.2529 [C 17 H 32 N] + as a highly sensitive marker signal (Scheme 1,d). [16,29,30,49] Isomeric structures were assigned based on the incorporation of branchedchain amino acids (BCAA).…”

Section: Resultsmentioning

confidence: 99%

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iso‐Fatty Acid Metabolism in Caenorhabditis elegans’ Ceramide Biosynthesis

Rivera Sánchez,

Bandi,

Scheidt

et al. 2023

Helvetica Chimica Acta

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Ceramide biosynthesis and its connection to iso‐fatty acid metabolism in the model organism Caenorhabditis elegans was investigated using a combination of reverse genetics and comparative ESI‐(+)‐HR‐MSe ceramide profiling along with incorporation experiments with bacterial mutants specifically enriched with isotopically labeled branched‐chain amino acids or branched‐chain fatty acids. Incorporation of a l‐leucine‐derived isovalerate unit into the conserved d17 : 1iso sphingosine building block proceeds through elo‐5 dependent chain elongation and depends on peroxisomal β‐oxidation by the 3‐ketoacyl‐CoA thiolase daf‐22, although ceramide profiles of N2 wildtype and daf‐22(ok693) are indistinguishable. Biosynthesis of the homologous N‐iso‐acyl moieties also depends on l‐leucine and isovalerate chain elongation but proceeds independently of elo‐5 and daf‐22. Biosynthesis of the dominating N‐docosanoyl moiety depends on elo‐3‐catalyzed chain elongation of bacteria‐derived palmitic acid, whereas the N‐tetracosanoyl moiety is derived from de novo lipogenesis.

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Section: Introductionmentioning

confidence: 75%

Section: Resultsmentioning

confidence: 99%

iso‐Fatty Acid Metabolism in Caenorhabditis elegans’ Ceramide Biosynthesis

Rivera Sánchez,

Bandi,

Scheidt

et al. 2023

Helvetica Chimica Acta

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“…The sphingolipids in C. elegans contain isomethyl branched sphingoid bases, which are biosynthesized from the condensation of 13-methyl myristoyl-CoA and serine . Previous analysis suggested that SPH id17:1 and SPH id17:0 were the dominant sphingoid bases in the sphingolipids of C. elegans . , Using the developed workflow shown in Figure a, we performed deep profiling of the total sphingoid bases in C. elegans . A total of 15 distinct structures of sphingoid bases were identified.…”

Section: Resultsmentioning

confidence: 99%

“…7 Previous analysis suggested that SPH id17:1 and SPH id17:0 were the dominant sphingoid bases in the sphingolipids of C. elegans. 41,42 Using the developed workflow shown in Figure 6a, we performed deep profiling of the total sphingoid bases in C. elegans. A total of 15 distinct structures of sphingoid bases were identified.…”

Section: Cid-triggered Rdd Of Tpn-derivatized Sphingoidmentioning

confidence: 99%

In-Depth Characterization of Sphingoid Bases via Radical-Directed Dissociation Tandem Mass Spectrometry

Zhao,

Qiao,

Xia

2023

J. Am. Soc. Mass Spectrom.

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Sphingoid base (SPH) is a basic structural unit of all classes of sphingolipids. A sphingoid base typically consists of an aliphatic chain that may be desaturated between C4 and C5, an amine group at C2, and a variable number of OH groups located at C1, C3, and C4. Variations in the chain length and the occurrence of chemical modifications, such as methyl branching, desaturation, and hydroxylation, lead to a large structural diversity and distinct functional properties of sphingoid bases. However, conventional tandem mass spectrometry (MS/MS) via collision-induced dissociation (CID) faces challenges in characterizing these modifications. Herein, we developed an MS/MS method based on CID-triggered radical-directed dissociation (RDD) for in-depth characterization of sphingoid bases. The method involves derivatizing the sphingoid amine with 3-(2,2,6,6-tetramethylpiperidin-1-yloxymethyl)-picolinic acid 2,5-dioxopyrrolidin-1-yl ester (TPN), followed by MS2 CID to unleash the pyridine methyl radical moiety for subsequent RDD. This MS/MS method was integrated on a reversed-phase liquid chromatography–mass spectrometry workflow and further applied for in-depth profiling of total sphingoid bases in bovine heart and Caenorhabditis elegans. Notably, we identified and relatively quantified a series of unusual sphingoid bases, including SPH id17:2 (4,13) and SPH it19:0 in C. elegans, revealing that the metabolic pathways of sphingolipids are more diverse than previously known.

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“…2D-RPLC × RPLC systems with high orthogonality [9][10] have been developed for the analysis of Curcuma kwangsiensis and Uncariae Ramulus Cum Unicis, respectively. Phenolic acids in Salvia miltiorrhiza [11], ginsenosides in Panax ginseng [12] and Panax notoginseng [13], sphingolipids in Caenorhabditis elegans [14], and proteins in tissues [15] have been comprehensively characterized using 2D-HILIC × RPLC, which demonstrate its enhanced separation efficiency, reduced ion suppression for the subsequent MS detection, and powerful characterization ability. Other orthogonal separation systems, including 2D-NPLC × RPLC [16], 2D-SEC × RPLC [17][18][19], 2D-chiral × chiral [20], and 2D-achiral × chiral [21], temperatureresponsive LC × RPLC [22], countercurrent chromatography × RPLC [23], IEC × assay-based RPLC [24], SFC × RPLC [25], IPC × RPLC [26], and IEC × SEC [27] have also been developed and applied toward the analysis of biosamples.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive profiling of Sanguisorba officinalis using offline two‐dimensional mixed‐mode liquid chromatography × reversed‐phase liquid chromatography, tandem high‐resolution mass spectrometry, and molecular network

Dai

Zhang

Xiong

et al. 2022

J of Separation Science

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The profiling of natural products is important in modern biological sciences and new drug development. However, the separation and characterization of complex herbal extracts are significantly challenging for researchers in the biochemical field. Herein, an offline two-dimensional mixed-mode liquid chromatography × reversed-phase liquid chromatography system is developed. Our system exhibits high orthogonality and is composed of a newly prepared stationary phase in the first dimension and a traditional C 18 phase in the second dimension, and is operated in combination with a high-resolution mass spectrometry and molecular network. Sanguisorba officinalis L. is studied using the proposed method owing to its bioactivity. With the aid of orthogonal separation, the ionization of the individual components is improved. The number of detected compounds and separated peaks are significantly increased when one-dimensional liquid chromatography is upgraded to two-dimensional liquid chromatography.In addition, 270 compounds (127 of which are tentatively characterized as new compounds, and further confirmation is needed) are successfully characterized based on their fragmentation patterns under the guidance of molecular network, while only 95 compounds are characterized using one-dimensional liquid chromatography and high-resolution mass spectrometry. The results indicate that the developed offline two-dimensional mixed-mode liquid chromatography ×

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Profiling of sphingolipids in Caenorhabditis elegans by two-dimensional multiple heart-cut liquid chromatography – mass spectrometry

Cited by 11 publications

References 45 publications

iso‐Fatty Acid Metabolism in Caenorhabditis elegans’ Ceramide Biosynthesis

iso‐Fatty Acid Metabolism in Caenorhabditis elegans’ Ceramide Biosynthesis

In-Depth Characterization of Sphingoid Bases via Radical-Directed Dissociation Tandem Mass Spectrometry

Comprehensive profiling of Sanguisorba officinalis using offline two‐dimensional mixed‐mode liquid chromatography × reversed‐phase liquid chromatography, tandem high‐resolution mass spectrometry, and molecular network

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