Quantifying the spectrum of DNA, RNA and protein adducts arising from lipid peroxidation in cells

TOXI 19

C. Eric Elmquist, elmquist@mit.edu1, Matthew R. Sullivan, matsul30@MIT.EDU1, Jose L. McFaline, jose_mc@mit.edu1, Jimmy Flarakos, flarakosj@alum.mit.edu1, Rosa G. Liberman1, Paul L Skipper, skipper@mit.edu2, Steven R. Tannenbaum2, and Peter C. Dedon, pcdedon@mit.edu1. (1) Biological Engineering Division, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Room NE47-293, Cambridge, MA 02139, (2) Biological Engineering Division and Department of Chemistry, Massachusetts Institute of Technology, Building 56, Room 753, 77 Massachusetts Ave., Cambridge, MA 02139
An emerging body of evidence points to lipid peroxidation as a major factor in the pathophysiology linking inflammation to human disease. We have developed a general approach to quantify lipid peroxidation-derived DNA, RNA and protein adducts. Membranes in human cells are labeled with 14C-containing polyunsaturated fatty acids (PUFA) and the cells are subjected to oxidative stress, followed by quantification of 14C-containing protein, DNA and RNA adducts by accelerator mass spectrometry (AMS). We have initiated these studies with human TK6 lymphoblastoid cells labeled with 14C-labeled linoleate, the most abundant PUFA in human cells. Following AMS quantification of total 14C in DNA and RNA fractions, individual RNA and DNA adducts were identified by nuclease digestion, HPLC resolution and AMS analysis of the fractions. This approach provides an opportunity to locate and characterize adducts containing a 14C label, which allows complete quantification and characterization of the spectrum of adducts arising from lipid peroxidation.
 

Young Investigator Session
8:15 AM-12:00 PM, Monday, August 20, 2007 BCEC -- 258C, Oral

Division of Chemical Toxicology

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007