Novel methods for the quantification of (2E)-hexadecenal by liquid chromatography with detection by either ESI QTOF tandem mass spectrometry or fluorescence measurement

Lüth, Anja; Neuber, Corinna; Kleuser, Burkhard

doi:10.1016/j.aca.2012.01.063

Cited by 15 publications

(17 citation statements)

References 30 publications

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“…Since FTY720-P was shown to lead to an internalization and degradation of S1PR 1 [42,43], we suggest that in those cells less S1PR 1 was present to mediate a S1P-dependent effect on DC maturation. Zeng et al postulated a FTY720-induced anergic phenotype switch of DCs, especially upon endotoxin activation [35]. Besides impaired phagocytotic, endocytotic and specific antigen presentation abilities observed in the FTY720-treated bone marrow-derived DCs (BM-DCs), they found a down-regulation of pro-inflammatory cytokines such as IL-6, TNF-α, IL-12p70 and MCP-1 in LPS-activated mature cells.…”

Section: S1p and Analogs Modify Development Of DC Subtypes From Commomentioning

confidence: 99%

“…Therefore, and to elucidate its role in physiological and pathological processes of DC homeostasis, a S1P lyase activity assay based on the determination of the S1P degradation product hexadecenal was established [35]. To this end, a high sensitivity such as mass spectrometry is required because only low levels of hexadecenal exist in biological samples such as serum, plasma and cells.…”

Section: S1p and Fingolimod Metabolism In Dendritic Cellsmentioning

confidence: 99%

“…Moreover, S1P lyase activity was modulated in cells by the use of 4-deoxypyridoxine, which is a competitive inhibitor of pyridoxal-5-phosphate, an essential cofactor of this enzyme. Mass spectrometry analysis after DAIH derivatisation revealed a significant decrease of hexadecenal compared to untreated cells, which allows the detection of S1P lyase activity also in DCs in response to immune modulatory stimuli [35]. …”

Section: S1p and Fingolimod Metabolism In Dendritic Cellsmentioning

confidence: 99%

“…To further understand the regulation of sphingolipid flow through the cell, the measurement of enzyme mRNA and protein (if possible at all), is not sufficient. Additionally, given the low frequency of blood or splenic DCs we started to optimize functional enzyme activity assays for SphK1 and SphK2 as well as S1P lyase [35]. In a first approach, we analyzed plain spleen cell suspensions, GM-CSF-/Flt3L-expanded, CD11c-enriched bone marrow cell preparations or DC/LC cell lines.…”

Section: S1p and Fingolimod Metabolism In Dendritic Cellsmentioning

confidence: 99%

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Sphingosine-1-Phosphate Modulates Dendritic Cell Function: Focus on Non-Migratory Effects in Vitro and in Vivo

Arlt

Schwiebs

Japtok

et al. 2014

Cell Physiol Biochem

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Dendritic cells (DCs) are the cutting edge in innate and adaptive immunity. The major functions of these antigen-presenting cells are the capture, endosomal processing and presentation of antigens, providing them an exclusive ability to provoke adaptive immune responses and to induce and control tolerance. Immature DCs capture and process antigens, migrate towards secondary lymphoid organs where they present antigens to naive T cells in a well-synchronized sequence of procedures referred to as maturation. Indeed, recent research indicated that sphingolipids are modulators of essential steps in DC homeostasis. It has been recognized that sphingolipids not only modulate the development of DC subtypes from precursor cells but also influence functional activities of DCs such as antigen capture, and cytokine profiling. Thus, it is not astonishing that sphingolipids and sphingolipid metabolism play a substantial role in inflammatory diseases that are modulated by DCs. Here we highlight the function of sphingosine 1-phosphate (S1P) on DC homeostasis and the role of S1P and S1P metabolism in inflammatory diseases.

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Section: S1p and Analogs Modify Development Of DC Subtypes From Commomentioning

confidence: 99%

Section: S1p and Fingolimod Metabolism In Dendritic Cellsmentioning

confidence: 99%

Section: S1p and Fingolimod Metabolism In Dendritic Cellsmentioning

confidence: 99%

Section: S1p and Fingolimod Metabolism In Dendritic Cellsmentioning

confidence: 99%

See 2 more Smart Citations

Sphingosine-1-Phosphate Modulates Dendritic Cell Function: Focus on Non-Migratory Effects in Vitro and in Vivo

Arlt

Schwiebs

Japtok

et al. 2014

Cell Physiol Biochem

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“…In addition, Jansen et al ( 15 ) used 2-hydroxyphytanoyl-CoA as a substrate for HACL1 and analyzed the produced pristanal as an ethoxime derivative by GC-MS. Related to SGPL1, other groups introduced alternative substrates which give rise to a fl uorescent aldehyde, either directly such as -NBD-sphingosine-1-phosphate ( 16 ) and -BODIPY-C 13 -sphingosine-1-phosphate ( 17 ), or via ␤ -elimination such as -(2-oxo-2H-chromen-7-yloxy)-C 5 -sphingosine-1-phosphate ( 18 ). Alternatively, SGPL1-produced aldehydes were detected by mass spectrometry after suitable derivatization with semicarbazide followed by LC-ESI (sphingosine-1-phosphate as substrate) ( 19 ), pentafl uorobenzyloxime followed by GC-EI (C 17 -sphinganine-1-phosphate as substrate) ( 20 ), or 2-diphenylacetyl-1,3-indandione-1-hydrazone followed by LC-MS/MS (sphingosine-1-phosphate as substrate) ( 21 ).…”

Section: Acidic Pretreatment Of Tissue Homogenatesmentioning

confidence: 99%

RP-HPLC-fluorescence analysis of aliphatic aldehydes: application to aldehyde-generating enzymes HACL1 and SGPL1

Mezzar

Schryver

Veldhoven

2014

Journal of Lipid Research

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Long-chain aliphatic aldehydes are produced during various metabolic processes, such as microsomal oxidation of long-chain alcohols, lysosomal degradation of prenylated proteins, peroxisomal ␣ -oxidation of 3-methylbranched fatty acids and 2-hydroxy long-chain fatty acids, microsomal breakdown of phosphorylated sphingoid bases, attack of plasmalogens by myeloperoxidase (MPO)-derived hypochlorous acid, and microsomal degradation of (lyso)plasmalogens (see Fig. 1A ). The involved enzymes are fatty aldehyde dehydrogenase (ALDH3A2) ( 1, 2 ), prenylcysteine oxidase 1 (PCYOX1) ( 3, 4 ), 2-hydroxyacylCoA lyase (HACL1) ( 5, 6 ), sphingosine-1-phosphate lyase (SGPL1) ( 7-9 ), MPO ( 10 ), and lysoplasmalogenase ( 11,12 ). In particular, two of these enzymes, HACL1 and SGPL1, have intensively been studied in our laboratory. Their activity measurements are complicated by their low specifi c activity [ ‫ف‬ 115 ( 5 ) and ‫ف‬ 15 mU/g rat liver ( 13 ), respectively], low K m (10-15 M), and the considerable interference of enzymes acting on their substrates (acylCoA hydrolases for HACL1; phosphatases for SGPL1) or on their cofactors (phosphatases acting on TPP, cofactor for HACL1, and on pyridoxal-phosphate, cofactor for SGPL1). In addition, the generated aldehyde is chemically not stable and subject to further metabolism, and compounds normally used to trap the aldehyde or block its metabolism interfere with the cofactor (pyridoxalphosphate). Moreover, the presence of plasmalogens in mammalian tissues, which give rise to aldehydes under acidic conditions ( 14 ), can cause a high background when using nonradioactive substrates, especially in nervous tissue.To measure HACL1 and SGPL1 activities in tissue or cell lysates, various protocols were developed by our group.Abstract Long-chain aldehydes are commonly produced in various processes, such as peroxisomal ␣ -oxidation of longchain 3-methyl-branched and 2-hydroxy fatty acids and microsomal breakdown of phosphorylated sphingoid bases. The enzymes involved in the aldehyde-generating steps of these processes are 2-hydroxyacyl-CoA lyase (HACL1) and sphingosine-1-phosphate lyase (SGPL1), respectively. In the present work, nonradioactive assays for these enzymes were developed employing the Hantzsch reaction. Tridecanal (C13-al) and heptadecanal (C17-al) were selected as model compounds and cyclohexane-1,3-dione as 1,3-diketone, and the fl uorescent derivatives were analyzed by reversed phase (RP)-HPLC. Assay mixture composition, as well as pH and heating, were optimized for C13-al and C17-al. Under optimized conditions, these aldehydes could be quantifi ed in picomolar range and different long-chain aldehyde derivatives were well resolved with a linear gradient elution by RP-HPLC. Aldehydes generated by recombinant enzymes could easily be detected via this method. Moreover, the assay allowed to document activity or defi ciency in tissue homogenates and fi broblast lysates without an extraction step. In conclusion, a simple, quick, and cheap assay for the study of HACL1 and SGPL1 ac...

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Mass Spectrometric Determination of Fatty Aldehydes Exemplified by Monitoring the Oxidative Degradation of (2E)-Hexadecenal in HepG2 Cell Lysates

et al. 2017

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Novel methods for the quantification of (2E)-hexadecenal by liquid chromatography with detection by either ESI QTOF tandem mass spectrometry or fluorescence measurement

Cited by 15 publications

References 30 publications

Sphingosine-1-Phosphate Modulates Dendritic Cell Function: Focus on Non-Migratory Effects <b><i>in Vitro</i></b> and <b><i>in Vivo</i></b>

Sphingosine-1-Phosphate Modulates Dendritic Cell Function: Focus on Non-Migratory Effects <b><i>in Vitro</i></b> and <b><i>in Vivo</i></b>

RP-HPLC-fluorescence analysis of aliphatic aldehydes: application to aldehyde-generating enzymes HACL1 and SGPL1

Mass Spectrometric Determination of Fatty Aldehydes Exemplified by Monitoring the Oxidative Degradation of (2E)-Hexadecenal in HepG2 Cell Lysates

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