Cross-linking and modification of fibronectin by peroxynitrous acid: Mapping and quantification of damage provides a new model for domain interactions.

Abstract

Fibronectin (FN) is an abundant glycoprotein found in plasma and the extracellular matrix (ECM). It is present at high concentrations at sites of tissue damage, where it is exposed to oxidants generated by activated leukocytes, including peroxynitrous acid (ONOOH) formed from nitric oxide (from inducible nitric oxide synthase) and superoxide radicals (from NADPH oxidases and other sources). ONOOH reacts rapidly with the abundant tyrosine and tryptophan residues in ECM proteins, resulting in the formation of 3-nitrotyrosine, di-tyrosine, and 6-nitrotryptophan. We have shown previously that human plasma FN is readily modified by ONOOH, but the extent and location of modifications, and the role of FN structure (compact versus extended) in determining these factors is poorly understood. Here, we provide a detailed LC-MS analysis of ONOOH-induced FN modifications, including the extent of their formation and the sites of intramolecular and intermolecular cross-links, including Tyr-Tyr, Trp-Trp, and Tyr-Trp linkages. The localization of these cross-links to specific domains provides novel data on the interactions between different modules in the compact conformation of plasma FN and allows us to propose a model of its unknown quaternary structure. Interestingly, the pattern of modifications is significantly different to that generated by another inflammatory oxidant, HOCl, in both extent and sites. The characterization and quantification of these modifications offers the possibility of the use of these materials as specific biomarkers of ECM modification and turnover in the many pathologies associated with inflammation-associated fibrosis.

• Authors: Michele Mariotti, Adelina Rogowska-Wrzesinska, Per Hägglund, Michael J Davies
• Journal: The Journal of biological chemistry (J Biol Chem) 2021 Jan-Jun Vol. 296 Pg. 100360 ISSN: 1083-351X [Electronic] United States
• PMID: 33539924 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.

Chemical References

• FN1 protein, human
• Fibronectins
• Oxidants
• Peroxynitrous Acid
• 3-nitrotyrosine
• Tyrosine
• 6-nitrotryptophan
• Tryptophan

Topics

• Atherosclerosis (metabolism)
• Cells, Cultured
• Chromatography, Gel (methods)
• Extracellular Matrix (metabolism, physiology)
• Fibronectins (chemistry, metabolism, physiology)
• Humans
• Inflammation (metabolism)
• Oxidants (metabolism)
• Oxidation-Reduction
• Peroxynitrous Acid (chemistry, pharmacology)
• Protein Domains (physiology)
• Tryptophan (analogs & derivatives, chemistry)
• Tyrosine (analogs & derivatives, chemistry)