Proteomic Identification of Carbonylated Proteins and Their Oxidation Sites

Abstract

Excessive oxidative stress leaves a protein carbonylation fingerprint in biological systems. Carbonylation is an irreversible post translational modification (PTM) that often leads to the loss of protein function and can be a component of multiple diseases. Protein carbonyl groups can be generated directly (by amino acids oxidation and the a-amidation pathway) or indirectly by forming adducts with lipid peroxidation products or glycation and advanced glycation end-products. Studies of oxidative stress are complicated by the low concentration of oxidation products and wide array of routes by which proteins are carbonylated. The development of new selection and enrichment techniques coupled with advances in mass spectrometry are allowing identification of hundreds of new carbonylated protein products from a broad range of proteins located at many sites in biological systems. The focus of this review is on the use of proteomics tools and methods to identify oxidized proteins along with specific sites of oxidative damage and the consequences of protein oxidation.

Chemical Isotope Labeling LC-MS for Monitoring Disease Progression and Treatment in Animal Models: Plasma Metabolomics Study of Osteoarthritis Rat Model

“…Protein oxidization is associated with many human diseases including cancer, neurodegenerative disorders and diabetes4142. Proteomics approach has now been widely used to search for new oxidative modifications in proteins, study the role of oxidization in proteins on cellular signal transduction pathways to understand disease pathogenesis and progression, and determine the usage of analyzing protein oxidization as potential biomarkers of human diseases434445. Direct analysis of 2-AAA, instead of analyzing oxidized proteins, has been used to determine 2-AAA as a biomarker for autoimmunity and age-associated changes in human collagen46.…”

Redox Proteomics in Selected Neurodegenerative Disorders: From Its Infancy to Future Applications

“…Nongel based strategies for the enrichment of PCO-modified proteins have been developed. A summary of these methods is provided in Figure 11 and has been recently reviewed (263). Most of these methods are based on the Schiff base chemistry that is possible with the carbonyl group.…”

“…(To see this illustration in color the reader is referred to the web version of this article at www.liebertonline.com/ars.) modification by lipid peroxidation products such as HNE and advanced glycation endproducts (AGEs) (263). It is often the case that other oxidative modifications such as oxidation of histidine, methionine, and tryptophan residues are also identified with these approaches.…”

Advances in purification and separation of posttranslationally modified proteins

“…These oxidations are believed to be irreversible and result in glutamic semialdehyde (arginine and proline), 2-amino-3-ketobutyric acid (threonine) or aminoadipic semialdehyde (lysine) (Fig. 5, G) [119][120][121][122]. Indirect protein carbonylation is more prevalent and involves a reaction with oxidized AGE (advanced glycation end) products (described in glycoproteomics, section 3.4), or hydroxyl radical-mediated oxidation of lipids.…”

“…Lipid-derived aldehydes can form Michael adducts with cysteine, histidine, lysine and arginine, and in some cases can further undergo Schiff base formation with amines of adjacent lysines, producing cross-linked amino acids [120,123,124]. Analysis of the resulting PTM diversity is hindered by their instability because carbonyl groups readily undergo Schiff base formation with proximate lysine residues, even when stored at -80 °C [122].…”