The rat leukocyte-type 12-lipoxygenase exhibits an intrinsic hepoxilin A3 synthase activity - PubMed (original) (raw)
. 2004 Jul 9;279(28):29023-30.
doi: 10.1074/jbc.M307576200. Epub 2004 May 3.
Affiliations
- PMID: 15123652
- DOI: 10.1074/jbc.M307576200
Free article
The rat leukocyte-type 12-lipoxygenase exhibits an intrinsic hepoxilin A3 synthase activity
Santosh Nigam et al. J Biol Chem. 2004.
Free article
Abstract
Hepoxilins are biologically relevant eicosanoids formed via the 12-lipoxygenase pathway of the arachidonic acid cascade. Although these eicosanoids exhibit a myriad of biological activities, their biosynthetic mechanism has not been investigated in detail. We examined the arachidonic acid metabolism of RINm5F rat insulinoma cells and found that they constitutively express a leukocyte-type 12S-lipoxygenase. Moreover, we observed that RINm5F cells exhibit an active hepoxilin A(3) synthase that converts exogenous 12S-HpETE (12S-5Z,8-Z,10E,14Z-12-hydro(pero)xy-eicosa-5,8,10,14-tetraenoic acid) or arachidonic acid predominantly to hepoxilin A(3). 12S-lipoxygenase and hepoxilin A(3) synthase activities were co-localized in the cytosol; immunoprecipitation with an anti-12S-lipoxygenase antibody co-precipitated the two catalytic activities. These data suggested that hepoxilin A(3) synthase activity may be considered an intrinsic catalytic property of the leukocyte-type 12S-lipoxygenase. To test this hypothesis we cloned the leukocyte-type 12S-LOX from RINm5F cells, expressed it in Pichia pastoris, and found that the recombinant enzyme exhibited both 12S-lipoxygenase and hepoxilin A(3) synthase activities. The recombinant human platelet-type 12S-lipoxygenase and the porcine leukocyte-type 12S-lipoxygenase also exhibited hepoxilin A(3) synthase activity. In contrast, the native rabbit reticulocyte-type 15S-lipoxygenase did not convert 12S-HpETE to hepoxilin isomers. These data suggest that the positional specificity of lipoxygenases may be crucial for this catalytic function. This hypothesis was confirmed by site-directed mutagenesis studies that altered the positional specificity of the rat leukocyte-type 12S- and the rabbit reticulocyte-type 15-lipoxygenase. In summary, it may be concluded that naturally occurring 12S-lipoxygenases exhibit an intrinsic hepoxilin A(3) synthase activity that is minimal in lipoxygenase isoforms with different positional specificity.
Similar articles
- Hepoxilin A3 synthase.
Nigam S, Zafiriou MP. Nigam S, et al. Biochem Biophys Res Commun. 2005 Dec 9;338(1):161-8. doi: 10.1016/j.bbrc.2005.09.065. Epub 2005 Sep 21. Biochem Biophys Res Commun. 2005. PMID: 16198304 Review. - Structure, biochemistry and biology of hepoxilins: an update.
Nigam S, Zafiriou MP, Deva R, Ciccoli R, Roux-Van der Merwe R. Nigam S, et al. FEBS J. 2007 Jul;274(14):3503-3512. doi: 10.1111/j.1742-4658.2007.05910.x. Epub 2007 Jul 2. FEBS J. 2007. PMID: 17608719 Review. - Structural basis for lipoxygenase specificity. Conversion of the human leukocyte 5-lipoxygenase to a 15-lipoxygenating enzyme species by site-directed mutagenesis.
Schwarz K, Walther M, Anton M, Gerth C, Feussner I, Kuhn H. Schwarz K, et al. J Biol Chem. 2001 Jan 5;276(1):773-9. doi: 10.1074/jbc.M005114200. J Biol Chem. 2001. PMID: 11027682 - Biosynthesis of hepoxilins: evidence for the presence of a hepoxilin synthase activity in rat insulinoma cells.
Shankaranarayanan P, Ciccoli R, Nigam S. Shankaranarayanan P, et al. FEBS Lett. 2003 Mar 13;538(1-3):107-12. doi: 10.1016/s0014-5793(03)00151-0. FEBS Lett. 2003. PMID: 12633862 - Alterations in leukotriene synthase activity of the human 5-lipoxygenase by site-directed mutagenesis affecting its positional specificity.
Schwarz K, Gerth C, Anton M, Kuhn H. Schwarz K, et al. Biochemistry. 2000 Nov 28;39(47):14515-21. doi: 10.1021/bi001447n. Biochemistry. 2000. PMID: 11087405
Cited by
- Discovering selective antiferroptotic inhibitors of the 15LOX/PEBP1 complex noninterfering with biosynthesis of lipid mediators.
Dar HH, Mikulska-Ruminska K, Tyurina YY, Luci DK, Yasgar A, Samovich SN, Kapralov AA, Souryavong AB, Tyurin VA, Amoscato AA, Epperly MW, Shurin GV, Standley M, Holman TR, St Croix CM, Watkins SC, VanDemark AP, Rana S, Zakharov AV, Simeonov A, Marugan J, Mallampalli RK, Wenzel SE, Greenberger JS, Rai G, Bayir H, Bahar I, Kagan VE. Dar HH, et al. Proc Natl Acad Sci U S A. 2023 Jun 20;120(25):e2218896120. doi: 10.1073/pnas.2218896120. Epub 2023 Jun 16. Proc Natl Acad Sci U S A. 2023. PMID: 37327313 Free PMC article. - Synthesis and Significance of Arachidonic Acid, a Substrate for Cyclooxygenases, Lipoxygenases, and Cytochrome P450 Pathways in the Tumorigenesis of Glioblastoma Multiforme, Including a Pan-Cancer Comparative Analysis.
Korbecki J, Rębacz-Maron E, Kupnicka P, Chlubek D, Baranowska-Bosiacka I. Korbecki J, et al. Cancers (Basel). 2023 Feb 2;15(3):946. doi: 10.3390/cancers15030946. Cancers (Basel). 2023. PMID: 36765904 Free PMC article. Review. - Clinical Significance of Lipid Transport Function of ABC Transporters in the Innate Immune System.
Kotlyarov S, Kotlyarova A. Kotlyarov S, et al. Membranes (Basel). 2022 Oct 31;12(11):1083. doi: 10.3390/membranes12111083. Membranes (Basel). 2022. PMID: 36363640 Free PMC article. Review. - The Biosynthesis of Enzymatically Oxidized Lipids.
Hajeyah AA, Griffiths WJ, Wang Y, Finch AJ, O'Donnell VB. Hajeyah AA, et al. Front Endocrinol (Lausanne). 2020 Nov 19;11:591819. doi: 10.3389/fendo.2020.591819. eCollection 2020. Front Endocrinol (Lausanne). 2020. PMID: 33329396 Free PMC article. Review. - Biotransformation of polyunsaturated fatty acids to bioactive hepoxilins and trioxilins by microbial enzymes.
An JU, Song YS, Kim KR, Ko YJ, Yoon DY, Oh DK. An JU, et al. Nat Commun. 2018 Jan 9;9(1):128. doi: 10.1038/s41467-017-02543-8. Nat Commun. 2018. PMID: 29317615 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases