Abstract
Glutathione peroxidase 3 (Gpx3) is ubiquitously expressed and is important antioxidant enzyme in yeast. It modulates the activities of redox-sensitive thiol proteins, particularly those involved in signal transduction pathway and protein translocation. Through immunoprecipitation/two-dimensional gel electrophoresis (IP-2DE), MALDI-TOF mass spectrometry, and a pull down assay, we found glutamine synthetase (GS; EC 6.3.1.2) as a candidate interacting protein with Gpx3. GS is a key enzyme in nitrogen metabolism and ammonium assimilation. It has been known that GS is non-enzymatically cleaved by ROS generated by MFO (thiol/ Fe3+/O2 mixed-function oxidase) system. In this study, it is demonstrated that GS interacts with Gpx3 through its catalytic domain both in vivo and in vitro regardless of redox state. In addition, Gpx3 helps to protect GS from inactivation and degradation via oxidative stress in an activity-independent manner. Based on the results, it is suggested that Gpx3 protects GS from non-enzymatic proteolysis, thereby contributing to cell homeostasis when cell is exposed to oxidative stress.
| Original language | English |
|---|---|
| Pages (from-to) | 405-409 |
| Number of pages | 5 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 362 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2007 Oct 19 |
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Keywords
- Glutamine synthetase
- Glutathione peroxidase 3
- MFO system
- Oxidative stress
- ROS
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
Glutathione peroxidase 3 of Saccharomyces cerevisiae suppresses non-enzymatic proteolysis of glutamine synthetase in an activity-independent manner. / Lee, Phil Young; Kho, Chang Won; Lee, Do Hee; Kang, Sunghyun; Kang, Seong Man; Lee, Sang Chul; Park, Byoung Chul; Cho, Sayeon; Bae, Kwang Hee; Park, Sung Goo.
In: Biochemical and Biophysical Research Communications, Vol. 362, No. 2, 19.10.2007, p. 405-409.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Glutathione peroxidase 3 of Saccharomyces cerevisiae suppresses non-enzymatic proteolysis of glutamine synthetase in an activity-independent manner
AU - Lee, Phil Young
AU - Kho, Chang Won
AU - Lee, Do Hee
AU - Kang, Sunghyun
AU - Kang, Seong Man
AU - Lee, Sang Chul
AU - Park, Byoung Chul
AU - Cho, Sayeon
AU - Bae, Kwang Hee
AU - Park, Sung Goo
PY - 2007/10/19
Y1 - 2007/10/19
N2 - Glutathione peroxidase 3 (Gpx3) is ubiquitously expressed and is important antioxidant enzyme in yeast. It modulates the activities of redox-sensitive thiol proteins, particularly those involved in signal transduction pathway and protein translocation. Through immunoprecipitation/two-dimensional gel electrophoresis (IP-2DE), MALDI-TOF mass spectrometry, and a pull down assay, we found glutamine synthetase (GS; EC 6.3.1.2) as a candidate interacting protein with Gpx3. GS is a key enzyme in nitrogen metabolism and ammonium assimilation. It has been known that GS is non-enzymatically cleaved by ROS generated by MFO (thiol/ Fe3+/O2 mixed-function oxidase) system. In this study, it is demonstrated that GS interacts with Gpx3 through its catalytic domain both in vivo and in vitro regardless of redox state. In addition, Gpx3 helps to protect GS from inactivation and degradation via oxidative stress in an activity-independent manner. Based on the results, it is suggested that Gpx3 protects GS from non-enzymatic proteolysis, thereby contributing to cell homeostasis when cell is exposed to oxidative stress.
AB - Glutathione peroxidase 3 (Gpx3) is ubiquitously expressed and is important antioxidant enzyme in yeast. It modulates the activities of redox-sensitive thiol proteins, particularly those involved in signal transduction pathway and protein translocation. Through immunoprecipitation/two-dimensional gel electrophoresis (IP-2DE), MALDI-TOF mass spectrometry, and a pull down assay, we found glutamine synthetase (GS; EC 6.3.1.2) as a candidate interacting protein with Gpx3. GS is a key enzyme in nitrogen metabolism and ammonium assimilation. It has been known that GS is non-enzymatically cleaved by ROS generated by MFO (thiol/ Fe3+/O2 mixed-function oxidase) system. In this study, it is demonstrated that GS interacts with Gpx3 through its catalytic domain both in vivo and in vitro regardless of redox state. In addition, Gpx3 helps to protect GS from inactivation and degradation via oxidative stress in an activity-independent manner. Based on the results, it is suggested that Gpx3 protects GS from non-enzymatic proteolysis, thereby contributing to cell homeostasis when cell is exposed to oxidative stress.
KW - Glutamine synthetase
KW - Glutathione peroxidase 3
KW - MFO system
KW - Oxidative stress
KW - ROS
UR - http://www.scopus.com/inward/record.url?scp=34548265133&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34548265133&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2007.08.035
DO - 10.1016/j.bbrc.2007.08.035
M3 - Article
C2 - 17707771
AN - SCOPUS:34548265133
VL - 362
SP - 405
EP - 409
JO - The BMJ
JF - The BMJ
SN - 0730-6512
IS - 2
ER -