title: Neuroprotectin D1/protectin D1 stereoselective and specific binding with human retinal pigment epithelial cells and neutrophils.
abstract: Retinal pigment epithelial (RPE) cells, derived from the neuroectoderm, biosynthesize the novel lipid mediator neuroprotectin D1 (NPD1) from docosahexaenoic acid (DHA) in response to oxidative stress or to neurotrophins, and in turn, elicits cytoprotection. Here, we report the identification of a 16,17-epoxide-containing intermediate in the biosynthesis of NPD1 in ARPE-19 cells from 17S-hydro-(peroxy)-docosahexaenoic acid. We prepared and isolated tritium-labeled NPD1 ([(3)H]-NPD1) and demonstrate specific and high-affinity stereoselective binding to ARPE-19 cells (K(d)=31.3+/-13.1 pmol/mg of cell protein). The stereospecific NPD1 interactions with these cells in turn gave potent protection against oxidative stress-induced apoptosis, and other structurally related compounds were weak competitors of NPD1 specific binding. This [(3)H]-NPD1/PD1 also displayed specific and selective high affinity binding with isolated human neutrophils (K(d) approximately 25 nM). Neither resolvin E1 nor lipoxin A(4) competed for [(3)H]-NPD1/PD1 specific binding with human neutrophils. Together, these results provide evidence for stereoselective specific binding of NPD1/PD1 with retinal pigment epithelial cells as well as human neutrophils. Moreover, they suggest specific receptors for this novel mediator in both the immune and visual systems.
authors.affiliation: Neuroscience Center of Excellence, Louisiana State University Health Sciences Center, School of Medicine, 2020 Gravier Street, Ste. D, New Orleans, LA 70112, USA.
authors.forename: Victor L
authors.initials: VL
authors.lastname: Marcheselli
authors.forename: Pranab K
authors.initials: PK
authors.lastname: Mukherjee
authors.forename: Makoto
authors.initials: M
authors.lastname: Arita
authors.forename: Song
authors.initials: S
authors.lastname: Hong
authors.forename: Rajee
authors.initials: R
authors.lastname: Antony
authors.forename: Kristopher
authors.initials: K
authors.lastname: Sheets
authors.forename: Jeremy W
authors.initials: JW
authors.lastname: Winkler
authors.forename: Nicos A
authors.initials: NA
authors.lastname: Petasis
authors.forename: Charles N
authors.initials: CN
authors.lastname: Serhan
authors.forename: Nicolas G
authors.initials: NG
authors.lastname: Bazan
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D000975:Antioxidants / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001667:Binding, Competitive; D002460:Cell Line; D004281:Docosahexaenoic Acids / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015118:Eicosapentaenoic Acid / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D004852:Epoxy Compounds / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D007536:Isomerism; D007700:Kinetics; D044045:Lipoxins / Q000378:metabolism; D018696:Neuroprotective Agents / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects; D055213:Retinal Pigment Epithelium / Q000187:drug effects / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000633:toxicity
chemical_list: C548872:17-hydro-(peroxy)-docosahexaenoic acid; D000894:Anti-Inflammatory Agents, Non-Steroidal; D000975:Antioxidants; D004852:Epoxy Compounds; D044045:Lipoxins; D018696:Neuroprotective Agents; D014409:Tumor Necrosis Factor-alpha; C040527:lipoxin A4; C487679:protectin D1; C518399:resolvin D1; D004281:Docosahexaenoic Acids; D015118:Eicosapentaenoic Acid; D006861:Hydrogen Peroxide; C499823:5S,12R,18R-trihydroxy-6Z,8E,10E,14Z,16E-eicosapentaenoic acid