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Oxidation of C18 Hydroxy-Polyunsaturated Fatty Acids to Epoxide or Ketone by Catalase-Related Hemoproteins Activated with Iodosylbenzene

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Lipids

Abstract

Small catalase-related hemoproteins with a facility to react with fatty acid hydroperoxides were examined for their potential mono-oxygenase activity when activated using iodosylbenzene. The proteins tested were a Fusarium graminearum 41 kD catalase hemoprotein (Fg-cat, gene FGSG_02217), a Pseudomonas fluorescens Pfl01 catalase (37.5 kD, accession number WP_011333788.1), and a Mycobacterium avium ssp. paratuberculosis 33 kD catalase (gene MAP-2744c). 13-Hydroxy-octadecenoic acids (which are normally unreactive) were selected as substrates because these enzymes react specifically with the corresponding 13S-hydroperoxides (Pakhomova et al. 18:2559–2568, 5; Teder et al. 1862:706–715, 14). In the presence of iodosylbenzene Fg-cat converted 13S-hydroxy-fatty acids to two products: the 15,16-double bond of 13S-hydroxy α-linolenic acid was oxidized stereospecifically to the 15S,16R-cis-epoxide or the 13-hydroxyl was oxidized to the 13-ketone. Products were identified by UV, HPLC, LC–MS, NMR and by comparison with authentic standards prepared for this study. The Pfl01-cat displayed similar activity. MAP-2744c oxidized 13S-hydroxy-linoleic acid to the 13-ketone, and epoxidized the double bonds to form the 9,10-epoxy-13-hydroxy, 11,12-epoxy-13-hydroxy, and 9,10-epoxy-13-keto derivatives; equivalent transformations occurred with 9S-hydroxy-linoleic acid as substrate. In parallel incubations in the presence of iodosylbenzene, human catalase displayed no activity towards 13S-hydroxy-linoleic acid, as expected from the highly restricted access to its active site. The results indicated that with suitable transformation to Compound I, monooxygenase activity can be demonstrated by these catalase-related hemoproteins with tyrosine as the proximal heme ligand.

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Abbreviations

PhIO:

Iodosylbenzene

cAOS:

Catalase-related allene oxide synthase

COSY:

Correlation spectroscopy

Fg-cat:

Fusarium graminearum catalase of the gene FGSG_02217

H(P)ODE:

Hydro(pero)xyoctadecadienoic acid

H(P)OTE:

Hydro(pero)xyoctadecatrienoic acid

HODE:

Hydroxyoctadecadienoic acid

LOX:

Lipoxygenase

MAP-2744c:

Mycobacterium avium ssp. paratuberculosis catalase of gene MAP-2744c

mCPBA:

Meta-chloroperoxybenzoic acid

Pfl01-cat:

Pseudomonas fluorescens Pf01 catalase from the gene, accession number WP_011333788.1

RP-HPLC:

Reversed phase high pressure liquid chromatography

SP-HPLC:

Straight phase high pressure liquid chromatography

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Acknowledgements

This work was supported in part by National Institutes of Health Grants GM-074888 and GM-15431 (to A. R. B.) and by the Institutional Research Funding IUT19-9 of the Estonian Ministry of Education and Research and the Estonian Science Foundation Grant 9410 (to Professor Nigulas Samel).

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  1. Tarvi Teder

    Present address: Department of Chemistry, Tallinn University of Technology, Tallinn, Estonia

Authors and Affiliations

  1. Department of Pharmacology and the Vanderbilt Institute of Chemical Biology, Vanderbilt, University School of Medicine, Nashville, TN, 37232, USA

    Tarvi Teder, William E. Boeglin & Alan R. Brash

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Correspondence to Alan R. Brash.

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Teder, T., Boeglin, W.E. & Brash, A.R. Oxidation of C18 Hydroxy-Polyunsaturated Fatty Acids to Epoxide or Ketone by Catalase-Related Hemoproteins Activated with Iodosylbenzene. Lipids 52, 587–597 (2017). https://doi.org/10.1007/s11745-017-4271-0

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