Confirmation of Multiple Organofluorophate Hydrolyzing Activities in the Protozoan Tetrahymena thermophila.
Abstract
The term DFPase describes an enzyme capable of hydrolyzing an organofluoromonophosphate. Partially purified preparations of DFPase were subjected to flatbed thin-layer isoelectric focusing (IEF) in polyacrylamide gels with an ampholine carrier of pH 4-6.5. Until recently, the separation of DFPases was completed on molecular sizing columns (sephacryl S-300).1 IEF provides information on the protein's isoelectric point. The isoelectric pH is the pH at which the net charge of the protein is zero. The use of IEF has confirmed the existence of multiple DFPases contained within the protozoan Tetrahymena thermophila. These experiments have confirmed the multiple DFPase entities as first discovered by sephacryl molecular sizing columns. Four areas of activity have been isolated. The next stage of research is to run the partially purified preparations of DFPase on a preparative flatbed electrofocusing in a granulated gel. This method would allow applying a maximum sample size of 4.5 ml. These gels will separate such a large volume that the eluded protein bands would have a concentration large enough to be applied to molecular sizing columns.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 1987
- Accession Number
- ADA188393
Entities
People
- Mark V. Haley
- Wayne G. Landis
Organizations
- Edgewood Chemical Biological Center