S-Nitrosylation Proteome Profile of Peripheral Blood Mononuclear Cells in Human Heart Failure

Abstract

Nitric oxide (NO) protects the heart against ischemic injury; however, NO- and superoxide-dependent S-nitrosylation (S-NO) of cysteines can affect function of target proteins and play a role in disease outcome. We employed 2D-GE with thiol-labeling FL-maleimide dye and MALDI-TOF MS/MS to capture the quantitative changes in abundance and S-NO proteome of HF patients (versus healthy controls, n = 30/group). We identified 93 differentially abundant (59-increased/34-decreased) and 111 S-NO-modified (63-increased/48-decreased) protein spots, respectively, in HF subjects (versus controls, fold-change | ≥1.5|, p ≤ 0.05). Ingenuity pathway analysis of proteome datasets suggested that the pathways involved in phagocytes' migration, free radical production, and cell death were activated and fatty acid metabolism was decreased in HF subjects. Multivariate adaptive regression splines modeling of datasets identified a panel of proteins that will provide >90% prediction success in classifying HF subjects. Proteomic profiling identified ATP-synthase, thrombospondin-1 (THBS1), and vinculin (VCL) as top differentially abundant and S-NO-modified proteins, and these proteins were verified by Western blotting and ELISA in different set of HF subjects. We conclude that differential abundance and S-NO modification of proteins serve as a mechanism in regulating cell viability and free radical production, and THBS1 and VCL evaluation will potentially be useful in the prediction of heart failure.

Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags

“…Supposable, the SNO proteome could be further preserved by use of appropriate chemical stabilizers or inhibitors additional to cooling and protease inhibitors, as it is common practice in example for phosphoproteomics. To our knowledge, such substances have not found widespread use yet, although some studies reported use of inhibitors ( Koo et al, 2016 , Foster et al, 2012 ). Lastly, there might be substantial species-derived differences between murine and human samples, which cannot be entirely distinguished from method-derived differences to this point.…”

Dysfunctional network and mutation genes of hypertrophic cardiomyopathy

“…Among the differentially expressed genes, the mRNA KRT1 with the largest upregulation multiple and the mRNA CYP1A1 with the largest down-regulation multiple may play an important role in the disease. Note that KRT1 expression increased in patients with heart failure and cardiac pressure overload (33,34). It has been reported that severe cytochrome P450 (P450) enzymes have been altered during cardiac hypertrophy (35).…”

Potential Role of Antioxidants as Adjunctive Therapy in Chagas Disease

“…In summary, a balance between the levels of ROS that are capable of inducing parasite damage and the antioxidant machinery that the host requires to detoxify and keep a safe environment for cells exposed to infection is fundamental. However, the host antioxidant response is exhausted during progression of Chagas disease [36,38,40,41] and a lack of appropriate antioxidant and repair response results in self-perpetuating mitochondrial dysfunction and ROS production in the heart (reviewed in [9,42]). The increased and sustained ROS production can signal the fibrotic gene expression and contribute to evolution of chronic cardiomyopathy [40,43].…”