Immunochemical detection of oxidized proteins formed by radiolysis of fractions of rat renal and hapatic tissue.
Keller-RJ; Bollinger-TL; Pumford-NR
Toxicologist 1995 Mar; 15(1):280
We have developed an immunochemical assay to detect carbonyl moieties that result from oxidative damage to proteins. Homogenates, microsomes and cytosol prepared from rat renal and hepatic tissue were exposed to hydroxyl radicals generated by a radiolysis mechanism. The resulting protein-derived carbonyls were reacted with 2,4-dinitrophenyl-hydrazine giving the corresponding hydrazones, which were detected by Western blot using anti-dinitrophenyl antisera. Analysis of the immunoblots using a densitometer indicated linear re-lationships between carbonyl group formation and increasing treatment from radiolysis in all fractions studied. In addition, distinct proteins stained more intensely in the immunoblots, particularly in the liver microsomes and cytosol, which demonstrated increased susceptibility of these proteins to oxidative damage. The characterization of susceptible proteins will increase our un-derstanding of the mechanisms of oxidative toxicity, and may prove useful as early indicators of oxidative protein damage.
Pulmonary-system-disorders; Analytical-methods; Immunochemistry; Carbonyls; Protein-chemistry; Blood-serum; Oxidative-processes; Analytical-processes
Pulmonary System Disorders
The Toxicologist. Society of Toxicology 34th Annual Meeting, March 5-9,1995, Baltimore, Maryland
University of Arkansas Med Scis Ltl Rock, Little Rock, Arkansas