2 research outputs found
Proteome profiling of s-nitrosylated synaptosomal proteins by isobaric mass tags.
peer reviewed• Protocol for quantitative proteomics of nitrosylation on synaptosomal proteins. • Identification of endogenous nitrosylation independent of induction by NO donors. • Use of iodoTMT sixplex mass tags for stable labeling, enrichment, identification, and multiplex quantitation. • Applicable on low amounts of sample material of mouse and human brain tissue.[Figure: see text
Quantitative proteomics of synaptosome S
peer reviewedIncreasing evidence suggests that both synaptic loss and neuroinflammation constitute early pathologic hallmarks of Alzheimer's disease. A downstream event during inflammatory activation of microglia and astrocytes is the induction of nitric oxide synthase type 2, resulting in an increased release of nitric oxide and the post-translational S-nitrosylation of protein cysteine residues. Both early events, inflammation and synaptic dysfunction, could be connected if this excess nitrosylation occurs on synaptic proteins. In the long term, such changes could provide new insight into patho-mechanisms as well as biomarker candidates from the early stages of disease progression. This study investigated S-nitrosylation in synaptosomal proteins isolated from APP/PS1 model mice in comparison to wild type and NOS2-/- mice, as well as human control, mild cognitive impairment and Alzheimer's disease brain tissues. Proteomics data were obtained using an established protocol utilizing an isobaric mass tag method, followed by nanocapillary high performance liquid chromatography tandem mass spectrometry. Statistical analysis identified the S-nitrosylation sites most likely derived from an increase in nitric oxide (NO) in dependence of presence of AD pathology, age and the key enzyme NOS2. The resulting list of candidate proteins is discussed considering function, previous findings in the context of neurodegeneration, and the potential for further validation studies