- 作者：Lena Dorothee Sch¨¹tte ; Stefan Baumeister ; Benjamin Weis ; Christoph Hudemann ; Eva-Maria Hanschmann ; Christopher Horst Lillig
- 关键词：Glutaredoxin ; Protein disulfide ; Intermediate trapping ; Proteomics ; Redox state ; Redox blot
- 刊名：Biochimica et Biophysica Acta (BBA)/General Subjects
- 出版年：2013
- 出版时间：November, 2013
- 年：2013
- 卷：1830
- 期：11
- 页码：4999-5005
- 全文大小：416 K
class=""h4"">Background
Glutaredoxins (Grxs) catalyze the reduction of protein disulfides via the dithiol mechanism and the de-/glutathionylation of substrates via the monothiol mechanism. These rapid, specific, and generally also reversible modifications are part of various signaling cascades regulating for instance cell proliferation, differentiation and apoptosis. Even though crucial functions of the conserved, mitochondrial Grx2a and the cytosolic/nuclear Grx2c isoforms have been proposed, only a few substrates have been identified in vitro or in vivo. The significance of redox signaling is emerging, yet a general lack of methods for the time-resolved analysis of these distinct and rapid modifications in vivo constitutes the biggest challenge in the redox signaling field.class=""h4"">Methods and results
Here, we have identified potential interaction partners for Grx2 isoforms in human HeLa cells and mouse tissues by an intermediate trapping approach. Some of the 50 potential substrates are part of the cytoskeleton or act in protein folding, cellular signaling and metabolism. Part of these interactions were further verified by immunoprecipitation or a newly established 2-D redox blot.
class=""h4"">Conclusions
Our study demonstrates that Grx2 catalyzes both the specific oxidation and the reduction of cysteinyl residues in the same compartment at the same time and without affecting the global cellular thiol-redox state.
class=""h4"">General significance
The knowledge of specific targets will be helpful in understanding the functions of Grx2. The 2-D redox blot may be useful for the analysis of the overall thiol-redox state of proteins with high molecular weight and numerous cysteinyl residues, that evaded analysis by previously described methods.