AUTHOR=Hisabori Toru , Hara Satoshi
TITLE=Kinetic analysis of the interactions between plant thioredoxin and target proteins
JOURNAL=Frontiers in Plant Science
VOLUME=4
YEAR=2013
URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2013.00508
DOI=10.3389/fpls.2013.00508
ISSN=1664-462X
ABSTRACT=
Thioredoxin is a critical protein that mediates the transfer of reducing equivalents in vivo and regulates redox sensitive enzymes in several cases. In addition, thioredoxin provides reducing equivalents to oxidoreductases such as peroxiredoxin. Through a dithiol–disulfide exchange reaction, the reduced form of thioredoxin preferentially interacts with the oxidized forms of targets, which are immediately released after this reaction is complete. In order to more thoroughly characterize these interactions between thioredoxin and its target proteins, a mutant version of thioredoxin that lacked the second cysteine was synthesized and interactions were monitored by surface plasmon resonance. The binding rates of thioredoxin to its targets were very different depending on the use of reducing equivalents by the targets: the enzymes whose activity was controlled by reduction or oxidation of a cysteine pair(s) in the molecule and the enzymes that used reducing equivalents provided by thioredoxin for their catalysis. In addition, thioredoxin revealed a stronger preference for an oxidized target. These results explain the reason for selective association of thioredoxin with oxidized targets for reduction, whereas immediate dissociation from a reduced target when the dithiol–disulfide exchange reaction is complete.