Effects of glutathione reductase inhibition on intracellular redox systems

BIOL 201

Yong Zhao, yzhao1788@jacks.sdstate.edu1, Teresa Seefeldt, Teresa.Seefeldt@sdstate.edu1, Wei Chen1, Xiuqing Wang2, Duane Matthees3, and Xiangming Guan, Xiangming.Guan@sdstate.edu1. (1) Department of Pharmaceutical Sciences, South Dakota State University, Brookings, SD 57007, (2) Department of Biology and Microbiology, (3) Department of Veterinary Science, South Dakota State University, Brookings, SD 57007, Brookings, SD 57007
Glutathione reductase (GR) is an important enzyme in maintaining an intracellular reducing environment, which is critical for the cell against oxidative stress. By employing a monkey kidney cell line and 2-acetylamino-3-[4-(2-acetylamino-2-carboxyethylsulfanylthiocarbonylamino)phenylthiocarbamoylsulfanyl]propionic acid (2-AAPA) - a selective irreversible GR inhibitor developed in this laboratory, we examined the effects of GR inhibition on intracellular reduced (GSH) and oxidized (GSSG) glutathione, thiols (-SH) and disulfides (-S-S-), glutathione peroxidase (GP), catalase, superoxide dismutase (SOD), gamma-glutamylcysteine synthetase (GCS), and glutathione synthetase (GS) over a 24 h period. GSH and GSSG were determined by LC/MS. Thiols and disulfides were quantified by HPLC. Western blot and real time RT-PCR were employed for the analysis of GP, GR, SOD, catalase, GCS, and GS. A significant increase in oxidative stress was observed when GR activity was inhibited by 90%. The experimental details and results will be presented.
 

Frontiers in Chemical Biology
5:00 PM-7:00 PM, Wednesday, August 22, 2007 BCEC -- Exhibit Hall - B2, Poster

Division of Biological Chemistry

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