da730d74ea92b858c5e29d18a0012c9d pmc_10383952.pdf a7076f6e1c435a385411af99fe2de890d5ffc9cf pmc_10383952.pdf 8202d8ff3b01112ae546aa9eaec05b01c5a7ed5d3790b02c7e027d26bace0c97 pmc_10383952.pdf Title: Different Structures—Similar Effect: Do Substituted 5-(4-Methoxyphenyl)-1H-indoles and 5-(4-Methoxyphenyl)-1H-imidazoles Represent a Common Pharmacophore for Substrate Selective Inhibition of Linoleate OxygenaseActivity of ALOX15? Subject: Mammalian 15-lipoxygenases (ALOX15) are lipid peroxidizing enzymes that exhibit variable functionality in different cancer and inflammation models. The pathophysiological role of linoleic acid- and arachidonic acid-derived ALOX15 metabolites rendered this enzyme a target for pharmacological research. Several indole and imidazole derivatives inhibit the catalytic activity of rabbit ALOX15 in a substrate-specific manner, but the molecular basis for this allosteric inhibition remains unclear. Here, we attempt to define a common pharmacophore, which is critical for this allosteric inhibition. We found that substituted imidazoles induce weaker inhibitory effects when compared with the indole derivatives. In silico docking studies and molecular dynamics simulations using a dimeric allosteric enzyme model, in which the inhibitor occupies the substrate-binding pocket of one monomer, whereas the substrate fatty acid is bound at the catalytic center of another monomer within the ALOX15 dimer, indicated that chemical modification of the core pharmacophore alters the enzyme–inhibitor interactions, inducing a reduced inhibitory potency. In our dimeric ALOX15 model, the structural differences induced by inhibitor binding are translated to the hydrophobic dimerization cluster and affect the structures of enzyme–substrate complexes. These data are of particular importance since substrate-specific inhibition may contribute to elucidation of the putative roles of ALOX15 metabolites derived from different polyunsaturated fatty acids inmammalian pathophysiology. Keywords: eicosanoids; lipoxygenase inhibitors; protein–protein interactions; allosteric inhibition; molecular dynamics Author: Alexander Zhuravlev, Alejandro Cruz, Vladislav Aksenov, Alexey Golovanov, José M. Lluch, Hartmut Kuhn, Àngels González-Lafont and Igor Ivanov Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sun Jul 16 02:37:04 2023 CEST ModDate: Sun Jul 16 02:40:12 2023 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 21 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 5054100 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 SBMXUO+VnURWPalladioL Type 1 Custom yes yes no 15 0 OCQPJP+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 UMXVRY+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 NINTFG+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 PHXIRD+URWPalladioL-BoldItal Type 1 Custom yes yes no 32 0 PUSVWS+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 50 0 WMLSQN+EURM10 Type 1 Builtin yes yes no 72 0 HHBSSZ+CMSY10 Type 1 Builtin yes yes no 79 0 XIRVNA+EURB10 Type 1 Builtin yes yes no 98 0 RFXLVC+SFRM1000 Type 1 Custom yes yes no 151 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-02-02 02:33:28 CET RepresentationInformation: pmc_10383952.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-09-16 02:17:11 CEST Size: 5054100 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 419 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: Chemical Synthesis of Target Compounds Destination: subsection.2.1 Item: Title: Inhibitory Potency of Substituted 4-Methoxyphenyl-1H-indole- and 4-Methoxyphenyl-1H-imidazol-Based Inhibitors 1 3 Destination: subsection.2.2 Item: Title: Substrate Selective Inhibition of LA-Oxygenase Activity of ALOX15 in the Presenceof Liposomes Destination: subsection.2.3 Item: Title: Docking and MD Simulations of Enzyme Inhibitor Complexes Destination: subsection.2.4 Item: Title: Impact of the Allosteric Inhibitors on Binding of LA and AA along Their MD Simulations Destination: subsection.2.5 Item: Title: Discussion Destination: section.3 Children: Item: Title: Liposomes Do Not Affect the Substrate Selective Inhibition of LA-Oxygenase Activityof ALOX15 Destination: subsection.3.1 Item: Title: Effect of the Inhibitor Structure on Cooperativity of Rabbit ALOX15 Monomers Destination: subsection.3.2 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Chemistry Destination: subsection.4.1 Item: Title: Biochemistry and Biotechnology Destination: subsection.4.2 Children: Item: Title: Expression of Rabbit ALOX15 in Bioreactor Destination: subsubsection.4.2.1 Item: Title: Enzyme Purification Destination: subsubsection.4.2.2 Item: Title: Liposome Preparations Destination: subsubsection.4.2.3 Item: Title: Rabbit ALOX15 Kinetic Assay Destination: subsubsection.4.2.4 Item: Title: Inhibitor Potency Assay Destination: subsubsection.4.2.5 Item: Title: Molecular Docking Studies Destination: subsubsection.4.2.6 Item: Title: Molecular Dynamics Simulations Destination: subsubsection.4.2.7 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.A. Info: Title: Different Structures Similar Effect: Do Substituted 5-(4-Methoxyphenyl)-1H-indoles and 5-(4-Methoxyphenyl)-1H-imidazoles Represent a Common Pharmacophore for Substrate Selective Inhibition of Linoleate OxygenaseActivity of ALOX15? Author: Alexander Zhuravlev, Alejandro Cruz, Vladislav Aksenov, Alexey Golovanov, José M. Lluch, Hartmut Kuhn, Àngels González-Lafont and Igor Ivanov Subject: Mammalian 15-lipoxygenases (ALOX15) are lipid peroxidizing enzymes that exhibit variable functionality in different cancer and inflammation models. The pathophysiological role of linoleic acid- and arachidonic acid-derived ALOX15 metabolites rendered this enzyme a target for pharmacological research. Several indole and imidazole derivatives inhibit the catalytic activity of rabbit ALOX15 in a substrate-specific manner, but the molecular basis for this allosteric inhibition remains unclear. Here, we attempt to define a common pharmacophore, which is critical for this allosteric inhibition. We found that substituted imidazoles induce weaker inhibitory effects when compared with the indole derivatives. In silico docking studies and molecular dynamics simulations using a dimeric allosteric enzyme model, in which the inhibitor occupies the substrate-binding pocket of one monomer, whereas the substrate fatty acid is bound at the catalytic center of another monomer within the ALOX15 dimer, indicated that chemical modification of the core pharmacophore alters the enzyme inhibitor interactions, inducing a reduced inhibitory potency. In our dimeric ALOX15 model, the structural differences induced by inhibitor binding are translated to the hydrophobic dimerization cluster and affect the structures of enzyme substrate complexes. These data are of particular importance since substrate-specific inhibition may contribute to elucidation of the putative roles of ALOX15 metabolites derived from different polyunsaturated fatty acids inmammalian pathophysiology. 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