title: Oxidative and nitrosative modifications of tropoelastin prevent elastic fiber assembly in vitro.
abstract: Elastic fibers are extracellular structures that provide stretch and recoil properties of tissues, such as lungs, arteries, and skin. Elastin is the predominant component of elastic fibers. Tropoelastin (TE), the precursor of elastin, is synthesized mainly during late fetal and early postnatal stages. The turnover of elastin in normal adult tissues is minimal. However, in several pathological conditions often associated with inflammation and oxidative stress, elastogenesis is re-initiated, but newly synthesized elastic fibers appear abnormal. We sought to determine the effects of reactive oxygen and nitrogen species (ROS/RNS) on the assembly of TE into elastic fibers. Immunoblot analyses showed that TE is oxidatively and nitrosatively modified by peroxynitrite (ONOO(-)) and hypochlorous acid (HOCl) and by activated monocytes and macrophages via release of ONOO(-) and HOCl. In an in vitro elastic fiber assembly model, oxidatively modified TE was unable to form elastic fibers. Oxidation of TE enhanced coacervation, an early step in elastic fiber assembly, but reduced cross-linking and interactions with other proteins required for elastic fiber assembly, including fibulin-4, fibulin-5, and fibrillin-2. These findings establish that ROS/RNS can modify TE and that these modifications affect the assembly of elastic fibers. Thus, we speculate that oxidative stress may contribute to the abnormal structure and function of elastic fibers in pathological conditions.
authors.affiliation: Departments of Medicine, Washington University School of Medicine, St Louis, Missouri 63110, USA.
authors.forename: Kamal
authors.initials: K
authors.lastname: Akhtar
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Broekelmann
authors.forename: Ming
authors.initials: M
authors.lastname: Miao
authors.forename: Fred W
authors.initials: FW
authors.lastname: Keeley
authors.forename: Barry C
authors.initials: BC
authors.lastname: Starcher
authors.forename: Richard A
authors.initials: RA
authors.lastname: Pierce
authors.forename: Robert P
authors.initials: RP
authors.lastname: Mecham
authors.forename: Tracy L
authors.initials: TL
authors.lastname: Adair-Kirk
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004547:Elastic Tissue / Q000378:metabolism*; D000071840:Fibrillin-2; D000071837:Fibrillins; D006801:Humans; D006997:Hypochlorous Acid / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D009000:Monocytes / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D030421:Peroxynitrous Acid / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D014333:Tropoelastin / Q000235:genetics / Q000378:metabolism*
chemical_list: C000605723:FBN2 protein, human; C000605728:Fbn2 protein, mouse; D000071840:Fibrillin-2; D000071837:Fibrillins; D008840:Microfilament Proteins; D014333:Tropoelastin; D030421:Peroxynitrous Acid; D006997:Hypochlorous Acid