Involvement of Electrostatic Interactions between the Components of Toluene Dioxygenase from Pseudomonas putida Fl

Abstract

A carboxyl group modifier, l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was used to study the interactions between three components of toluene dioxygenase (TDO) from Pseudomonas putida FI. $Ferredoxin_{TOL}$ activity was increased by the treatment with EDC; however, the activity was rapidly declined in the prolonged incubation. In covalent cross-linking experiments with EDC, $Ferredoxin_{TOL}$ made a one-to-one complex with $Reductase_{TOL}$ or the large subunit of $ISP_{TOL}$. These results provide evidence for the involvement of electrostatic interactions in the TDO electron transfer system.

Keywords

• Pseudomonas;
• toluene dioxygenase;
• protein- protein interaction;
• l-ethyl-3-(3-dimethylaminopropyl) carbodiimide;
• cross-linking