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TretyakovaNY

Research Abstracts Online
January - December 2011

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University of Minnesota Twin Cities
Academic Health Center
Masonic Cancer Center

PI: Natalia Y. Tretyakova

Proteomics of DNA-Protein Cross-Links

1,2,3,4-Diepoxybutane (DEB) is a strongly genotoxic diepoxide hypothesized to be the ultimate carcinogenic metabolite of the common industrial chemical and environmental carcinogen 1,3-butadiene. DEB is a bis-electrophile capable of cross-linking cellular biomolecules to form DNA-DNA and DNA-protein cross-links (DPCs), which are thought to play a central role in its biological activity. Previous studies with recombinant proteins have shown that the biological outcomes of DEB-induced DPCs are strongly influenced by protein identities. This present study employs mass spectrometry-based proteomics and immunological detection to identify the proteins that form DPCs in cell culture. Following DNA isolation from treated and control cells (in triplicate) and the removal of non-covalently bound proteins, the cross-linked proteins are separated by gel electrophoresis and identified by bottom-up proteomics analysis.

Group Members

Thakshila D. Dissanayake Rallage, Graduate Student
Teshome Gherezghiher, Research Associate
Delshanee Kotandeniya, Research Associate
Xun Ming, Graduate Student
Darrin M. York, Fellow, Faculty Collaborator