title: Generation of Adducts of 4-Hydroxy-2-nonenal with Heat Shock 60 kDa Protein 1 in Human Promyelocytic HL-60 and Monocytic THP-1 Cell Lines.
abstract: Heat shock 60 kDa protein 1 (HSP60) is a chaperone and stress response protein responsible for protein folding and delivery of endogenous peptides to antigen-presenting cells and also a target of autoimmunity implicated in the pathogenesis of atherosclerosis. By two-dimensional electrophoresis and mass spectrometry, we found that exposure of human promyelocytic HL-60 cells to a nontoxic concentration (10 μM) of 4-hydroxy-2-nonenal (HNE) yielded a HSP60 modified with HNE. We also detected adducts of HNE with putative uncharacterized protein CXorf49, the product of an open reading frame identified in various cell and tissue proteomes. Moreover, exposure of human monocytic THP-1 cells differentiated with phorbol 12-myristate 13-acetate to 10 μM HNE, and to light density lipoprotein modified with HNE (HNE-LDL) or by copper-catalyzed oxidation (oxLDL), but not to native LDL, stimulated the formation of HNE adducts with HSP60, as detected by immunoprecipitation and western blot, well over basal levels. The identification of HNE-HSP60 adducts outlines a framework of mutually reinforcing interactions between endothelial cell stressors, like oxLDL and HSP60, whose possible outcomes, such as the amplification of endothelial dysfunction, the spreading of lipoxidative damage to other proteins, such as CXorf49, the activation of antigen-presenting cells, and the breaking of tolerance to HSP60 are discussed.
authors.affiliation: Dipartimento di Medicina e Scienze della Salute, Università del Molise, 86100 Campobasso, Italy.
authors.forename: Alessia
authors.identifier: 0000-0002-5924-3694
authors.initials: A
authors.lastname: Arcaro
authors.affiliation: Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, 10125 Torino, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Daga
authors.affiliation: Dipartimento di Medicina e Scienze della Salute, Università del Molise, 86100 Campobasso, Italy.
authors.forename: Giovanni Paolo
authors.initials: GP
authors.lastname: Cetrangolo
authors.affiliation: Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, 10125 Torino, Italy.
authors.forename: Eric Stefano
authors.initials: ES
authors.lastname: Ciamporcero
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, 80131 Napoli, Italy.
authors.forename: Alessio
authors.identifier: 0000-0002-2037-673X
authors.initials: A
authors.lastname: Lepore
authors.affiliation: Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, 10125 Torino, Italy.
authors.forename: Stefania
authors.identifier: 0000-0001-6937-8632
authors.initials: S
authors.lastname: Pizzimenti
authors.affiliation: Dipartimento di Medicina e Scienze della Salute, Università del Molise, 86100 Campobasso, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Petrella
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, 80131 Napoli, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Graf
authors.affiliation: Graduate School of Bioagricultural Science, Nagoya University, Nagoya 464-8601, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Uchida
authors.affiliation: Centro di Spettrometria di Massa Proteomica e Biomolecolare, ISA-CNR, 83100 Avellino, Italy.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Mamone
authors.affiliation: Dipartimento di Agraria, Università di Napoli "Federico II", Portici, 80055 Napoli, Italy.
authors.forename: Pasquale
authors.initials: P
authors.lastname: Ferranti
authors.affiliation: William Harvey Research Institute, Queen Mary University London, London E1 4NS, UK.
authors.forename: Paul R J
authors.initials: PR
authors.lastname: Ames
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, 80131 Napoli, Italy.
authors.forename: Giuseppe
authors.identifier: 0000-0001-9380-8436
authors.initials: G
authors.lastname: Palumbo
authors.affiliation: Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, 10125 Torino, Italy.
authors.forename: Giuseppina
authors.identifier: 0000-0002-8587-4166
authors.initials: G
authors.lastname: Barrera
authors.affiliation: Dipartimento di Medicina e Scienze della Salute, Università del Molise, 86100 Campobasso, Italy.
authors.forename: Fabrizio
authors.identifier: 0000-0002-6704-3115
authors.initials: F
authors.lastname: Gentile
mesh_terms: D000447:Aldehydes / Q000494:pharmacology*; D015153:Blotting, Western; D018834:Chaperonin 60 / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D018922:HL-60 Cells; D006801:Humans; D047468:Immunoprecipitation; D008856:Microscopy, Fluorescence; D024101:Mitochondrial Proteins / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000447:Aldehydes; D018834:Chaperonin 60; C060293:HSPD1 protein, human; D024101:Mitochondrial Proteins; D020543:Proteome; C027576:4-hydroxy-2-nonenal