8a39c38de0f6b86c36c49c05037eb2c4 pmc_10094721.pdf b093692eb1606afdcdeeb3ef63e2e844833c5b70 pmc_10094721.pdf 262d0ce7c5107d41e8e6335ae2090f29b2ae639444ffc94620e1d576c7704193 pmc_10094721.pdf Title: Hydroperoxidation of Docosahexaenoic Acid by Human ALOX12 and pigALOX15-mini-LOX Subject: Human lipoxygenase 12 (hALOX12) catalyzes the conversion of docosahexaenoic acid (DHA) into mainly 14S-hydroperoxy-4Z,7Z,10Z,12E,16Z,19Z-docosahexaenoic acid (14S-H(p)DHA). This hydroperoxidation reaction is followed by an epoxidation and hydrolysis process that finally leads to maresin 1 (MaR1), a potent bioactive specialized pro-resolving mediator (SPM) in chronic inflammation resolution. By combining docking, molecular dynamics simulations, and quantum mechanics/molecular mechanics calculations, we have computed the potential energy profile of DHA hydroperoxidation in the active site of hALOX12. Our results describe the structural evolution of the molecular system at each step of this catalytic reaction pathway. Noteworthy, the required stereospecificity of the reaction leading to MaR1 is explained by the configurations adopted by DHA bound to hALOX12, along with the stereochemistry of the pentadienyl radical formed after the first step of the mechanism. In pig lipoxygenase 15 (pigALOX15-mini-LOX), our calculations suggest that 14S-H(p)DHA can be formed, but with a stereochemistry that is inadequate for MaR1 biosynthesis. Keywords: hydroperoxidation mechanism; human platelet ALOX12; enzyme catalysis; molecular dynamics simulations; QM/MM calculations Author: Miquel Canyelles-Niño, Àngels González-Lafont and José M. Lluch Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Mar 23 10:34:08 2023 CET ModDate: Thu Mar 23 10:53:11 2023 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 29 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 21064095 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- EPTQDJ+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 MWINDN+VnURWPalladioL Type 1 Custom yes yes no 15 0 UKRFGI+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 JTJYUI+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 WJHTVE+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 IVXOCJ+CMSY10 Type 1 Builtin yes yes no 32 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 50 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 53 0 SQXHQP+EURM10 Type 1 Builtin yes yes no 75 0 HDABPA+PalatinoLinotype,Bold 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HDACAD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 599 0 HDADCP+PalatinoLinotype TrueType MacRoman yes yes no 602 0 HDABPA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 624 0 HDABPB+PalatinoLinotype CID TrueType Identity-H yes yes yes 627 0 HDABPC+PalatinoLinotype TrueType WinAnsi yes yes no 633 0 HDACAD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 636 0 HDADCP+PalatinoLinotype TrueType MacRoman yes yes no 639 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-02-02 02:35:38 CET RepresentationInformation: pmc_10094721.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-09-05 02:22:26 CEST Size: 21064095 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 931 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results and Discussion Destination: section.2 Children: Item: Title: MD Simulations of the DHA/hALOX12 Complex Destination: subsection.2.1 Item: Title: QM/MM Calculations of DHA Hydroperoxidation Catalyzed by hALOX12 Destination: subsection.2.2 Children: Item: Title: Hydrogen Abstraction Reactions Catalyzed by hALOX12 Destination: subsubsection.2.2.1 Item: Title: O2 Addition and Retro-Hydrogen Abstraction Reactions Catalyzed by hALOX12 Destination: subsubsection.2.2.2 Item: Title: MD Simulations of the DHA/pigALOX15-mini-LOX Complex Destination: subsection.2.3 Item: Title: QM/MM Calculations of H12-Abstraction from DHA Catalyzed by pigALOX15-mini-LOX Destination: subsection.2.4 Item: Title: Materials and Methods Destination: section.3 Children: Item: Title: Protein Setup Destination: subsection.3.1 Item: Title: Molecular Docking Simulations Destination: subsection.3.2 Item: Title: Molecular Dynamics Simulations Destination: subsection.3.3 Item: Title: QM/MM Calculations Destination: subsection.3.4 Item: Title: Conclusions Destination: section.4 Item: Title: References Destination: section.4 Info: Title: Hydroperoxidation of Docosahexaenoic Acid by Human ALOX12 and pigALOX15-mini-LOX Author: Miquel Canyelles-Niño, Àngels González-Lafont and José M. Lluch Subject: Human lipoxygenase 12 (hALOX12) catalyzes the conversion of docosahexaenoic acid (DHA) into mainly 14S-hydroperoxy-4Z,7Z,10Z,12E,16Z,19Z-docosahexaenoic acid (14S-H(p)DHA). This hydroperoxidation reaction is followed by an epoxidation and hydrolysis process that finally leads to maresin 1 (MaR1), a potent bioactive specialized pro-resolving mediator (SPM) in chronic inflammation resolution. By combining docking, molecular dynamics simulations, and quantum mechanics/molecular mechanics calculations, we have computed the potential energy profile of DHA hydroperoxidation in the active site of hALOX12. Our results describe the structural evolution of the molecular system at each step of this catalytic reaction pathway. Noteworthy, the required stereospecificity of the reaction leading to MaR1 is explained by the configurations adopted by DHA bound to hALOX12, along with the stereochemistry of the pentadienyl radical formed after the first step of the mechanism. In pig lipoxygenase 15 (pigALOX15-mini-LOX), our calculations suggest that 14S-H(p)DHA can be formed, but with a stereochemistry that is inadequate for MaR1 biosynthesis. 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