Functional domains of pseudomonas exotoxin identified by deletion analysis of the gene expressed in E. coli

Jaulang Hwang, David J. Fitzgerald, Sankar Adhya, Ira Pastan

Research output: Contribution to journalArticle

326 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)129-136
Number of pages8
JournalCell
Volume48
Issue number1
DOIs
Publication statusPublished - Jan 16 1987
Externally publishedYes

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ADP-Ribosylation Factors
Peptide Elongation Factor 2
Immunotoxins
Exotoxins
Poisons
Gene Deletion
Eukaryotic Cells
Pseudomonas
Adenosine Diphosphate
Escherichia coli
Genes
Membranes
Molecules
Proteins
Protein Domains
Pseudomonas aeruginosa toxA protein

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Functional domains of pseudomonas exotoxin identified by deletion analysis of the gene expressed in E. coli. / Hwang, Jaulang; Fitzgerald, David J.; Adhya, Sankar; Pastan, Ira.

In: Cell, Vol. 48, No. 1, 16.01.1987, p. 129-136.

Research output: Contribution to journalArticle

Hwang, Jaulang ; Fitzgerald, David J. ; Adhya, Sankar ; Pastan, Ira. / Functional domains of pseudomonas exotoxin identified by deletion analysis of the gene expressed in E. coli. In: Cell. 1987 ; Vol. 48, No. 1. pp. 129-136.
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