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Oxidative Medicine and Cellular Longevity
Volume 2015 (2015), Article ID 296146, 13 pages
Research Article

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

1Dipartimento di Medicina e Scienze della Salute, Università del Molise, 86100 Campobasso, Italy
2Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, 10125 Torino, Italy
3Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, 80131 Napoli, Italy
4Graduate School of Bioagricultural Science, Nagoya University, Nagoya 464-8601, Japan
5Centro di Spettrometria di Massa Proteomica e Biomolecolare, ISA-CNR, 83100 Avellino, Italy
6Dipartimento di Agraria, Università di Napoli “Federico II”, Portici, 80055 Napoli, Italy
7William Harvey Research Institute, Queen Mary University London, London E1 4NS, UK

Received 8 December 2014; Revised 20 April 2015; Accepted 29 April 2015

Academic Editor: Francisco Javier Romero

Copyright © 2015 Alessia Arcaro et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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.