September 7, 2016

Protein oxidation in cardiomyocyte model of OS

“Oxidative protein modifications caused by reactive oxygen and nitrogen species in cell cultures and rat models of hypoxia and nitrosative stress”

Funded by German Research Consul (Deutsche Forschungsgemeinschaft , DFG)

 

Druck

 

Cellular response to oxidative stress (OS) requires coordination of multiple regulatory pathways to adjust cellular behaviors and maintain homeostasis. Those regulations occur on different levels of cellular organization and involve changes in gene expression, protein synthesis and degradation rates, redox and signaling post-translational protein modifications, regulation of lipidome and metabolome. In order to understand the mechanisms OS response and propose targeted therapeutics for OS-related diseases, it is important to obtain an integrated view on cellular system response. Modern high-throughput analytical methods, such as mass spectrometry (MS) based omics techniques, are capable to provide qualitative and quantitative information at each level of cell organization. Further implementation of systems biology algorithms allows to integrate large number of omics-derived data to explain and predict systems behavior.
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We are working with a dynamic model of nitrosative/oxidative stress using rat primary cardiomyocytes (CM) treated with peroxynitrite donor (SIN-1) for 15, 30, 70 min, and 16 h to monitor the early and delayed effects of OS on cellular proteome, transriptome and lipidome. A large number of qualitative and quantitative MS-omics protocols are developed and applied to characterize system behavior. Thus we are using high resolution mass spectrometry for relative quantification of:

 

  • Phospholipids abundances
  • Changes in intracellular protein abundances
  • Cardiomyocyte secretome
  • Phospholipid oxidation
  • Nitrated and oxidized fatty acids
  • Oxysterols
  • Protein carbonylation
  • Protein-lipid adducts
  • Modification of Cys residues (oxidation, nitrosylation, glutathionylation)
  • Protein phosphorylation
  • Modification of tyrosine residues
All Omics data are integrated via systems biology tools and combined with biochemical assays to confirm predicted functional effects.

 

Researches involved in the project: