title: Identification of glutathione adducts of α-chlorofatty aldehydes produced in activated neutrophils.
abstract: α-Chlorofatty aldehydes (α-ClFALDs) are produced by hypochlorous acid targeting plasmalogens during neutrophil activation. This study investigated the reaction of the α-chlorinated carbon of α-ClFALD with the nucleophile, GSH. Utilizing ESI/MS/MS, the reaction product of GSH and the 16-carbon α-ClFALD, 2-chlorohexadecanal (2-ClHDA), was characterized. The resulting conjugate of 2-ClHDA and GSH (HDA-GSH) has an intact free aldehyde, and the chlorine at the α-carbon is ejected. Stable isotope-labeled [d4]HDA-GSH was synthesized, which further confirmed the structure, and was used to quantify natural α-ClFALD conjugates of GSH (FALD-GSH) using reverse-phase LC with detection by ESI/MS/MS using selected reaction monitoring. HDA-GSH is elevated in RAW 264.7 cells treated with physiologically relevant concentrations of exogenous 2-ClHDA. Furthermore, PMA-treated primary human neutrophils have elevated levels of HDA-GSH and the conjugate of 2-chlorooctadecanal (2-ClODA) and GSH (ODA-GSH), as well as elevated levels of 2-ClHDA and 2-ClODA. Production of both conjugates in PMA-stimulated neutrophils was reduced by 3-aminotriazole pretreatment, which also blocks endogenous α-ClFALD production. Additionally, plasma FALD-GSH levels were elevated in the K/BxN mouse arthritis model. Taken together, these studies demonstrate novel peptidoaldehydes derived from GSH and α-ClFALD in activated human neutrophils and in vivo in K/BxN mice.
authors: Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, MO 63104 Center for Cardiovascular Research, Saint Louis University School of Medicine, St. Louis, MO 63104.
authors: Mark A
authors: MA
authors: Duerr
authors: Department of Microbiology and Molecular Immunology, Saint Louis University School of Medicine, St. Louis, MO 63104.
authors: Rajeev
authors: R
authors: Aurora
authors: Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, MO 63104 Center for Cardiovascular Research, Saint Louis University School of Medicine, St. Louis, MO 63104.
authors: David A
authors: DA
authors: Ford
mesh_terms: D000447:Aldehydes / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D005227:Fatty Acids / Q000097:blood* / Q000737:chemistry; D005978:Glutathione / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D051379:Mice; D018375:Neutrophil Activation; D009504:Neutrophils / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: C432398:2-chlorohexadecanal; D000447:Aldehydes; D005227:Fatty Acids; D005978:Glutathione
keywords: fatty aldehydes; hypochlorous acid; myeloperoxidase; plasmalogen