Pseudomonas exotoxin A is a single chain toxin with three structural domains that inhibits protein synthesis in eukaryotic cells by catalyzing ADP ribosylation of elongation factor 2. To study the function of these domains, we deleted different portions of the PE structural gene and expressed these constructs in E. coli using an inducible T7 promoter. These studies indicate that structural domain la is required for cell recognition, that structural domain ll is required to translocate the toxin across a cellular membrane, and that structural domain III and a portion of domain lb are required for ADP ribosylation activity. Toxin lacking domain la is about 100-fold less toxic to mice than intact PE and should be a useful molecule for the construction of immunotoxins.
ASJC Scopus subject areas
- 生物化學、遺傳與分子生物學 (全部)