A. DNA Ligase III. DNA Ligase is a specific type of enzyme that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. It plays a role in repairing single-strand breaks in duplex DNA in living organisms. This protein has been investigated in different cells by SDS-PAGE coupled to mass spectrometric analyses. B. Telomerase. Telomerase is a ribonucleoprotein that is an enzyme that adds DNA sequence repeats to the 3'end of DNA strands. Induction of this protein was investigated by SDS-PAGE coupled to mass spectrometric analyses. C. Pulegone. Pulegone is a commonly-used flavoring agent (mint flavoring) and is known to be toxic in high doses. Using microscopy, it has been shown that glomerulopathy can occur when rats are given high doses of this chemical. To address the composition of the kidney of affected rats, we are using mass spectrometry to identify the proteins present as well as determine which proteins are affected by this chemical. This study should provide a significant amount of information that can help in understanding the mechanism of this chemical.

Project Start
Project End
Budget Start
Budget End
Support Year
21
Fiscal Year
2013
Total Cost
$120,054
Indirect Cost
City
State
Country
Zip Code
Schorzman, Allison N; Perera, Lalith; Cutalo-Patterson, Jenny M et al. (2011) Modeling of the DNA-binding site of yeast Pms1 by mass spectrometry. DNA Repair (Amst) 10:454-65
Hager-Braun, Christine; Hochleitner, Elisabeth O; Gorny, Miroslaw K et al. (2010) Characterization of a discontinuous epitope of the HIV envelope protein gp120 recognized by a human monoclonal antibody using chemical modification and mass spectrometric analysis. J Am Soc Mass Spectrom 21:1687-98
Dhungana, Suraj; Fessler, Michael B; Tomer, Kenneth B (2009) Epitope mapping by differential chemical modification of antigens. Methods Mol Biol 524:119-34
Hobbs, Carey A; Deterding, Leesa J; Perera, Lalith et al. (2009) Structural characterization of the conformational change in calbindin-D28k upon calcium binding using differential surface modification analyzed by mass spectrometry. Biochemistry 48:8603-14
Dhungana, Suraj; Williams, Jason G; Fessler, Michael B et al. (2009) Epitope mapping by proteolysis of antigen-antibody complexes. Methods Mol Biol 524:87-101
Chattopadhyay, Rima; Iacob, Roxana; Sen, Shalmali et al. (2009) Functional and structural characterization of factor Xa dimer in solution. Biophys J 96:974-86
Smedley 3rd, James G; Sharp, Joshua S; Kuhn, Jeffrey F et al. (2008) Probing the pH-dependent prepore to pore transition of Bacillus anthracis protective antigen with differential oxidative protein footprinting. Biochemistry 47:10694-704