Fluorescence labeling of carbonylated lipids and proteins in cells using coumarin-hydrazide

摘要: Carbonylation is a generic term which refers to reactive carbonyl groups present in biomolecules due oxidative reactions induced by oxygen species. Carbonylated proteins, lipids and nucleic acids have been intensively studied often associated with onset or progression of stress related disorders. In order reveal underlying carbonylation pathways biological relevance, it crucial study their intracellular formation spatial distribution. species are usually identified quantified cell lysates body fluids after derivatization using specific chemical probes. However, cellular tissue distribution less investigated. Here, we report coumarin-hydrazide, fluorescent probe for time- cost-efficient labeling carbonyls followed fluorescence microscopy evaluate both time space. The specificity coumarin-hydrazide was confirmed dose-dependent experiments human primary fibroblasts stressed paraquat compared conventional DNPH-based immunocytochemistry. Both techniques stained carbonylated accumulated cytoplasm strong perinuclear clustering. Using complimentary array analytical methods towards protein- lipid-bound has shown. Additionally, co-distribution oxidized phospholipids demonstrated.