Browsing by Author "McDonagh, Brian"
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- Ubiquitination and carbonylation as markers of oxidative-stress in Ruditapes decussatusPublication . Chora, Suze; McDonagh, Brian; Sheehan, David; Starita-Geribaldi, Mireille; Roméo, Michèle; Bebianno, Maria JoãoEnvironmental pollutants, such as metals, are widespread in aquatic environments and can lead to the formation of reactive oxygen species (ROS). ROS are highly toxic in marine species since they can cause serious reversible and irreversible changes in proteins including ubiquitination and modifications such as carbonylation. This study aimed to confirm the potential of ubiquitination and carbonylation as markers of oxidative stress in the clam Ruditapes decussatus (Veneroida, Veneridae) exposed to cadmium (40 microg/L). After 21 days of exposure clams were dissected into gills and digestive gland. Cytosolic proteins of both tissues were separated by two-dimensional electrophoresis (2-D SDS-PAGE) and analysed by immunobloting. Higher ubiquitination and carbonylation levels were in digestive gland of contaminated organisms. These results confirm the potential of ubiquitination and carbonylation as a sensitive and specific marker of oxidative stress in marine bivalves. In this approach, changes in protein structure provide options for affinity selection of sub-proteomes for 2D SDS-PAGE, simplifying the detection of protein biomarkers using proteomic approach.
- Ubiquitination and carbonylation of proteins in the clam Ruditapes decussatus, exposed to nonylphenol using redox proteomicsPublication . Chora, Suze; McDonagh, Brian; Sheehan, David; Starita-Geribaldi, Mireille; Roméo, Michèle; Bebianno, Maria JoãoUbiquitination and carbonylation of proteins were investigated in the gill and digestive gland of Ruditapes decussatus exposed to NP (nonylphenol) (100 μgL(-1)) using redox proteomics. After 21 d of exposure, clams were dissected and cytosolic proteins of both tissues separated by 2DE SDS-PAGE. Protein expression profiles were tissue-dependent and differently affected by NP exposure. Ubiquitination and carbonylation were also tissue-specific. NP exposure induced significantly more ubiquitinated proteins in gills than in digestive glands, compared to controls. Digestive gland showed a significant higher number of carbonylated proteins than gills after NP exposure. Protein ubiquitination and carbonylation are therefore independent processes. Results showed that NP exposure generated ROS in gill and digestive gland of R. decussatus that significantly altered the proteome. Results also highlighted the advantage of using redox proteomics in the assessment of protein ubiquitination and carbonylation, which may be markers of oxidative stress in R. decussatus.