Protein adduction by reactive lipid oxidation-derived electrophiles

TOXI 101

Simona G. Codreanu, simona.codreanu@vanderbilt.edu1, Bing Zhang, bing.zhang@vanderbilt.edu2, Jeremy S. Myers1, Scott M. Sobecki, scott.sobecki@vanderbilt.edu3, Dean D. Billheimer, dean.billheimer@vanderbilt.edu4, and D. C. Liebler1. (1) Department of Biochemistry, Vanderbilt University Medical Center, 465 21st Ave South, Nashville, TN 37232, (2) Department of Biomedical Informatics, Vanderbilt University Medical Center, 465 21st Ave South, Nashville, TN 37232, (3) Mass Spectrometry Research Center, Vanderbilt University Medical Center, 465 21st Ave South, Nashville, TN 37232, (4) Department of Biostatistics, Vanderbilt University Medical Center, 465 21st Ave South, Nashville, TN 37232
Lipid peroxidation products are thought to be mediators of oxidative stress in many environmentally-related diseases, but little is known about protein targets of these electrophiles. We characterized the protein targets of HNE, a prototypical lipid oxidation product. RKO cells were treated with increasing concentrations of HNE (0, 50, 100 µM) and adducted proteins were derivatized by reaction with biotinamidohexanoic acid hydrazide and captured with streptavidin. Affinity-captured proteins were analyzed by shotgun LC-MS-MS of tryptic peptides from 1D SDS-PAGE-separated proteins on a Thermo LTQ ion trap instrument. Peptides identified by database searching at 95% confidence were assembled to generate parsimonious protein lists. Totals of 320, 1173, and 1424 discernible proteins were identified for the untreated control, the 50 µM and 100 µM HNE-treated samples, respectively. Characterization of protein targets of lipid oxidation-derived electrophiles enables analysis of networks and signaling pathways impacted by endogenous electrophiles in oxidative stress.
 

Poster Session and Awards
6:00 PM-10:00 PM, Tuesday, August 21, 2007 BCEC -- 204 A/B, Poster

Division of Chemical Toxicology

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