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Protein Carbonyl Determination Using Biotin Hydrazide

  • Kenneth Hensley
  • Kelly S. Williamson
Part of the Methods in Pharmacology and Toxicology book series (MIPT)

Abstract

Protein oxidation is a recognized component of aging and a consequence of severe or prolonged exposure to reactive oxygen species (ROS). Direct attack of protein by ROS causes formation of protein-bound carbonyl groups (1). These carbonyl functions represent a variety of site-specific modifications, most particularly adipic and glutamic acid semialdehydes (2). Additionally, numerous lipid oxidation products, including αβ-unsaturated γ-hydroxyalkenals can attack proteins yielding protein-bound aldehydes (3). Furthermore, nonemzymatic glycation can yield protein-bound carbonyl functionalities (4). Thus, protein carbonyls represent a possibly convenient indicator of oxidative stress. A variety of techniques have been introduced to measure protein carbonyls in tissue extracts, where they are found to increase exponentially as a function of organism aging (5). All the extant techniques for protein carbonyl determination rely upon reductive amination between the carbonyl group and a probe, typically dinitrophenylhydrazine (DNPH) (5, 6). Antibodies specific to the probe can then be used to visualize protein carbonyls.

Keywords

Protein Carbonyl Reductive Amination Protein Carbonyl Level Carbonyl Function Organism Aging 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc.,Totowa, NJ 2003

Authors and Affiliations

  • Kenneth Hensley
    • 1
  • Kelly S. Williamson
    • 1
  1. 1.Free Radical Biology and Aging ProgramOklahoma Medical Research FoundationOklahoma City

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