Disulfide proteome in the analysis of protein function and structure

Abstract

Many proteins undergo post-translational modification via well defined mechanisms such as acetylation, phosphorylation and glycosylation and thereby control a spectrum of biochemical processes. A growing body of evidence suggests that the reversible reduction of disulfide bonds also alters the structure and activity of proteins. Thioredoxin, a ubiquitous 12 kDa protein with a catalytically active disulfide active site (Cys-Gly-Pro-Cys), plays a central role in controlling the redox status of disulfide bonds in proteins that regulate a range of processes. Included are photosynthesis, seed germination, transcription, cell division, radical scavenging and detoxification. The ability to identify unknown functions of proteins of all types has been advanced by the emerging field of functional proteomics. In this brief review, we introduce the disulfide proteome as a tool that complements other methods for the comprehensive analysis of proteins. In so doing, the usefulness of applying this method for both in vitro and in vivo analyses is discussed for thioredoxin and other disulfide proteins, especially those occurring in plants.

A Comprehensive Analysis of the Peroxiredoxin Reduction System in the Cyanobacterium Synechocystis sp. Strain PCC 6803 Reveals that All Five Peroxiredoxins Are Thioredoxin Dependent

“…1A and B). This would be consistent with the presence of intramolecular disulfides, which render the proteins more compact (74).…”

The Effect of α-Amanitin on the Arabidopsis Seed Proteome Highlights the Distinct Roles of Stored and Neosynthesized mRNAs during Germination

“…Transgenic tobacco plants expressing a dominant-negative form of RSG exhibit reduced germination frequency (Fukazawa et al, 2000), showing that RSG does play a role in seed germination. Recent work also documented the role played by thioredoxins, which are small M r proteins containing a redox-active disulfide group (Yano et al, 2002), in seed germination. This study showed that overexpression of wheat thioredoxin h in barley was associated with enhanced germination and increased GA levels in the transgenic germinating seeds (Wong et al, 2002), suggesting that thioredoxins can somehow regulate some of the enzymes in the GA biosynthetic pathway and/or factors modulating the rate and extent of GA biosynthesis.…”

The diversity and complexity of the cyanobacterial thioredoxin systems

“…To this end, several ingenious high-throughput screening procedures were developed. One method involves comparative labelling of protein thiols with the fluorescent compound monobromobimane (mBBr) before and after reduction by Trx (Yano et al 2001(Yano et al , 2002. Another approach takes advantage of the catalytic mechanism of Trx, which includes a short-lived mixed disulphide intermediate between the first cysteine of the Trx consensus active site (WCGPC) and a cysteine of the target protein (Wynn et al 1995).…”