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Biosynthesis of 14,15-Hepoxilins in Human L1236 Hodgkin Lymphoma Cells and Eosinophils

  • Original Article
  • Published:
Lipids

Abstract

Hepoxilins are epoxy alcohols synthesized through the 12-lipoxygenase (12-LO) pathway in animal cells. The epidermis is the principal source of hepoxilins in humans. Here we report on the formation of novel hepoxilin regioisomers formed by the 15-LO pathway in human cells. The Hodgkin lymphoma cell line L1236 possesses high 15-lipoxygenase-1 (15-LO-1) activity and incubation of L1236 cells with arachidonic acid led to the formation of 11(S)-hydroxy-14(S),15(S)-epoxy 5(Z),8(Z),12(E) eicosatrienoic acid (14,15-HxA3 11(S)) and 13(R)-hydroxy-14(S),15(S)-epoxy 5(Z),8(Z),11(Z) eicosatrienoic acid (14,15-HxB3 13 (R)). In addition, two hitherto unidentified products were detected and these products were collected and analyzed by positive ion electrospray tandem mass spectrometry. These metabolites were identified as 11(S),15(S)-dihydroxy-14(R)-glutathionyl-5(Z),8(Z),12(E)-eicosatrienoic acid (14,15-HxA3-C) and 11(S),15(S)-dihydroxy-14(R)-cysteinyl-glycyl-5(Z),8(Z),12(E)-eicosatrienoic acid (14,15-HxA3-D). Incubation of L1236 cells with synthetic 14,15-HxA3 11(S) also led to the formation of 14,15-HxA3-C and 14,15-HxA3-D. Several soluble glutathione transferases, in particular GST M1-1 and GST P1-1, were found to catalyze the conversion of 14,15-HxA3 to 14,15-HxA3-C. L1236 cells produced approximately twice as much eoxins as cysteinyl-containing hepoxilins upon stimulation with arachidonic acid. Human eosinophils, nasal polyps and dendritic cells selectively formed 14,15-HxA3 11(S) and 14,15-HxB3 13(R) stereoisomers, but not cysteinyl-containing hepoxilins, after stimulation with arachidonic acid. Furthermore, purified recombinant 15-LO-1 alone catalyzed the conversion of arachidonic acid to 14,15-HxA3 11(S) and 14,15-HxB3 13(R), showing that human 15-LO-1 possesses intrinsic 14,15-hepoxilin synthase activity.

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Abbreviations

EX:

Eoxin

GST:

Glutathione transferase

HPETE:

Hydroperoxyeicosatetraenoic acid

HPLC:

High performance liquid chromatography

Hx:

Hepoxilin

LC–MS:

Liquid chromatography–mass spectrometry

LO:

Lipoxygenase

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Acknowledgments

This work was supported by grants from the Karolinska Institutet, the Swedish Research Council, the Swedish Cancer Society, Orexo AB, and European Commission Sixth Framework Programme Grant LSHM-CT-2004-005033.

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Authors and Affiliations

  1. Department of Medicine, Karolinska Institutet and Karolinska University Hospital Solna, 171 76, Stockholm, Sweden

    Åsa Brunnström & Hans-Erik Claesson

  2. Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77, Stockholm, Sweden

    Mats Hamberg

  3. Institute of Mass Spectrometry, School of Medicine, Swansea University, Singleton Park, Swansea, SA2 8PP, UK

    William J. Griffiths

  4. Department of Biochemistry and Organic Chemistry, Uppsala University, Biomedical Center, 75123, Uppsala, Sweden

    Bengt Mannervik

  5. Department of Neurochemistry, Stockholms University, 106 91, Stockholm, Sweden

    Bengt Mannervik

  6. Orexo AB, Box 303, 75105, Uppsala, Sweden

    Åsa Brunnström & Hans-Erik Claesson

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  1. Åsa Brunnström
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  2. Mats Hamberg
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  3. William J. Griffiths
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  4. Bengt Mannervik
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  5. Hans-Erik Claesson
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Correspondence to Hans-Erik Claesson.

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Brunnström, Å., Hamberg, M., Griffiths, W.J. et al. Biosynthesis of 14,15-Hepoxilins in Human L1236 Hodgkin Lymphoma Cells and Eosinophils. Lipids 46, 69–79 (2011). https://doi.org/10.1007/s11745-010-3485-1

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  • DOI: https://doi.org/10.1007/s11745-010-3485-1

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