Affinity chromatography reveals RuBisCO as an ecdysteroid-binding protein.
The aim of this work was to isolate plant ecdysteroid-binding proteins using affinity chromatography. Ecdysteroids as insect hormones have been investigated thoroughly but their function and the mechanism of action in plants and other organisms is still unknown although ecdysteroids occur in some plants in a relatively large amount. Therefore, 20-hydroxyecdysone was immobilized on a polymeric carrier as a ligand for affinity chromatography in order to isolate plant ecdysteroid-binding proteins from the cytosolic extract of New Zealand spinach (Tetragonia tetragonoides). Non-specifically bound proteins were eluted with a rising gradient of concentration of sodium chloride, and 3% (v/v) acetic acid was used for the elution of the specifically bound proteins. Using this method, ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) was isolated. The influence of ecdysteroids on RuBisCO was further studied. Our results show that ecdysteroids are able to increase the yield of RuBisCO-mediated reaction in which CO(2) is fixed into organic matter by more than 10%.
Uhlik
Ondrej
O
Institute of Organic Chemistry and Biochemistry AS CR, Czech Academy of Sciences, Flemingovo n. 2, 166 10 Prague 6, Czech Republic.
Kamlar
Marek
M
Kohout
Ladislav
L
Jezek
Rudolf
R
Harmatha
Juraj
J
Macek
Tomas
T