title: On the cellular metabolism of the click chemistry probe 19-alkyne arachidonic acid.
abstract: Alkyne and azide analogs of natural compounds that can be coupled to sensitive tags by click chemistry are powerful tools to study biological processes. Arachidonic acid (AA) is a FA precursor to biologically active compounds. 19-Alkyne-AA (AA-alk) is a sensitive clickable AA analog; however, its use as a surrogate to study AA metabolism requires further evaluation. In this study, AA-alk metabolism was compared with that of AA in human cells. Jurkat cell uptake of AA was 2-fold greater than that of AA-alk, but significantly more AA-Alk was elongated to 22:4. AA and AA-alk incorporation into and remodeling between phospholipid (PL) classes was identical indicating equivalent CoA-independent AA-PL remodeling. Platelets stimulated in the pre-sence of AA-alk synthesized significantly less 12-lipoxygenase (12-LOX) and cyclooxygenase products than in the presence of AA. Ionophore-stimulated neutrophils produced significantly more 5-LOX products in the presence of AA-alk than AA. Neutrophils stimulated with only exogenous AA-alk produced significantly less 5-LOX products compared with AA, and leukotriene B4(LTB4)-alk was 12-fold less potent at stimulating neutrophil migration than LTB4, collectively indicative of weaker leukotriene B4receptor 1 agonist activity of LTB4-alk. Overall, these results suggest that the use of AA-alk as a surrogate for the study of AA metabolism should be carried out with caution.
authors.affiliation: Département de Chimie et Biochimie, Université de Moncton, Moncton, NB E1A 3E9, Canada; Centre de Recherche du Centre Hospitalier Universitaire de Québec, Département de Microbiologie et Immunologie, Université Laval, Québec, QC G1V 4G2, Canada.
authors.forename: Philippe Pierre
authors.initials: PP
authors.lastname: Robichaud
authors.affiliation: Département de Chimie et Biochimie, Université de Moncton, Moncton, NB E1A 3E9, Canada; Centre de recherche de l'Institut universitaire de cardiologie et de pneumologie de Québec, Université Laval, Québec, QC G1V 4G5, Canada.
authors.forename: Samuel J
authors.initials: SJ
authors.lastname: Poirier
authors.affiliation: Département de Chimie et Biochimie, Université de Moncton, Moncton, NB E1A 3E9, Canada.
authors.forename: Luc H
authors.initials: LH
authors.lastname: Boudreau
authors.affiliation: Département de Chimie et Biochimie, Université de Moncton, Moncton, NB E1A 3E9, Canada.
authors.forename: Jérémie A
authors.initials: JA
authors.lastname: Doiron
authors.affiliation: Atlantic Cancer Research Institute, Moncton, NB E1C 8X3, Canada.
authors.forename: David A
authors.initials: DA
authors.lastname: Barnett
authors.affiliation: Centre de Recherche du Centre Hospitalier Universitaire de Québec, Département de Microbiologie et Immunologie, Université Laval, Québec, QC G1V 4G2, Canada.
authors.forename: Eric
authors.initials: E
authors.lastname: Boilard
authors.affiliation: Département de Chimie et Biochimie, Université de Moncton, Moncton, NB E1A 3E9, Canada. Electronic address: marc.surette@umoncton.ca.
authors.forename: Marc E
authors.initials: ME
authors.lastname: Surette
mesh_terms: D001092:Arachidonate 12-Lipoxygenase / Q000378:metabolism*; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D001095:Arachidonic Acids / Q000138:chemical synthesis / Q000493:pharmacokinetics / Q000494:pharmacology; D057930:Click Chemistry; D006801:Humans; D019169:Jurkat Cells; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*
chemical_list: D001095:Arachidonic Acids; D010743:Phospholipids; D001092:Arachidonate 12-Lipoxygenase; D001094:Arachidonate 5-Lipoxygenase
keywords: cyclooxygenase; eicosanoid; leukotriene; lipoxygenase; phospholipid remodeling