title: Carbonylated plasma proteins as potential biomarkers of obesity induced type 2 diabetes mellitus.
abstract: Protein carbonylation is a common nonenzymatic oxidative post-translational modification, which is often considered as biomarker of oxidative stress. Recent evidence links protein carbonylation also to obesity and type 2 diabetes mellitus (T2DM), though the protein targets of carbonylation in human plasma have not been identified. In this study, we profiled carbonylated proteins in plasma samples obtained from lean individuals and obese patients with or without T2DM. The plasma samples were digested with trypsin, carbonyl groups were derivatized with O-(biotinylcarbazoylmethyl)hydroxylamine, enriched by avidin affinity chromatography, and analyzed by RPC-MS/MS. Signals of potentially modified peptides were targeted in a second LC-MS/MS analysis to retrieve the peptide sequence and the modified residues. A total of 158 unique carbonylated proteins were identified, of which 52 were detected in plasma samples of all three groups. Interestingly, 36 carbonylated proteins were detected only in obese patients with T2DM, whereas 18 were detected in both nondiabetic groups. The carbonylated proteins originated mostly from liver, plasma, platelet, and endothelium. Functionally, they were mainly involved in cell adhesion, signaling, angiogenesis, and cytoskeletal remodeling. Among the identified carbonylated proteins were several candidates, such as VEGFR-2, MMP-1, argin, MKK4, and compliment C5, already connected before to diabetes, obesity and metabolic diseases.
authors.affiliation: Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, ‡Center for Biotechnology and Biomedicine, and §Department of Medicine, Universität Leipzig , Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Ravi Chand
authors.initials: RC
authors.lastname: Bollineni
authors.forename: Maria
authors.initials: M
authors.lastname: Fedorova
authors.forename: Matthias
authors.initials: M
authors.lastname: Blüher
authors.forename: Ralf
authors.initials: R
authors.lastname: Hoffmann
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000097:blood* / Q000378:metabolism; D001798:Blood Proteins / Q000032:analysis* / Q000378:metabolism*; D016022:Case-Control Studies; D003924:Diabetes Mellitus, Type 2 / Q000097:blood* / Q000209:etiology; D005260:Female; D006801:Humans; D056865:Ideal Body Weight; D048670:MAP Kinase Kinase 4 / Q000097:blood / Q000378:metabolism; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000097:blood / Q000378:metabolism; D008875:Middle Aged; D009765:Obesity / Q000097:blood* / Q000150:complications; D050050:Protein Carbonylation; D040901:Proteomics / Q000379:methods; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000097:blood / Q000378:metabolism; D057188:Workflow
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D040301:Vascular Endothelial Growth Factor Receptor-2; D048670:MAP Kinase Kinase 4; C482559:MAP2K4 protein, human; D020781:Matrix Metalloproteinase 1
keywords: LC−MS/MS; aldehyde reactive probe (ARP); obesity; protein carbonylation; type 2 diabetes mellitus (T2DM)