An egf-pseudomonas exotoxin a recombinant protein with a deletion in toxin binding domain specifically kills egf receptor bearing cells

Chi Hon Lee, E. Ching Lee, Shih Tzer Tsai, Hsing Jien Kung, Yin Chang Liu, Jaulang Hwang

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We constructed two chimeric toxins; one composed of epidermal growth factor (EGF) and pseudomonas exotoxin A (PE), designated EGF-PE and the other composed of EGF and PE with a deletion of the Ia domain (cell-binding domain), designated EGF-PE (δIa). Both chimeric toxins reacted with anti-EGF and anti-PE antibodies. The cell-killing experiments showed that EGF-PE, but not EGF-PE(δIa), was cytotoxic to the murine fibroblast cell line NR6, which carried the PE receptor, but not the EGF receptor. However, after NR6 was transfected with DNA for the expression of human EGF receptor, the transfected cell line, designated NRHER5, overexpressed human EGF receptors and became sensitive to EGF-PE(δIA). The cytotoxicity of EGF-PE(δIa), but not EGF-PE, to NRHER5 can be completely blocked by an excess amount of EGF. To completely reverse the cytotoxicity of EGF-PE on NRHER5, both the EGF receptor pathway and the PE receptor pathway need to be blocked. These results suggest that EGF-PE exhibits both EGF and PE binding activities, while EGF-PE(δIA) possesses only EGF binding activity. Thus, EGF-PE(δIa) may be a better chimeric toxin than EGF-PE in terms of target specificity to EGF receptor bearing cells. We, therefore, examined the cytotoxicity of EGF-PE(δIa) to various human cancer cell lines. We find that human cancer cells containing more EGF receptors are more sensitive to EGF-PE(δIa).

Original languageEnglish
Pages (from-to)433-440
Number of pages8
JournalProtein Engineering, Design and Selection
Volume6
Issue number4
DOIs
Publication statusPublished - Jun 1993
Externally publishedYes

Fingerprint

Bearings (structural)
Exotoxins
Recombinant proteins
Pseudomonas
Recombinant Proteins
Epidermal Growth Factor
Epidermal Growth Factor Receptor
Immunotoxins
Cytotoxicity
Cells
Pseudomonas aeruginosa toxA protein
Cell Line

Keywords

  • Chimera
  • EGF
  • Pseudomonas exotoxin A

ASJC Scopus subject areas

  • Pharmacology
  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Molecular Biology
  • Bioengineering
  • Biotechnology
  • Biochemistry

Cite this

An egf-pseudomonas exotoxin a recombinant protein with a deletion in toxin binding domain specifically kills egf receptor bearing cells. / Lee, Chi Hon; Lee, E. Ching; Tsai, Shih Tzer; Kung, Hsing Jien; Liu, Yin Chang; Hwang, Jaulang.

In: Protein Engineering, Design and Selection, Vol. 6, No. 4, 06.1993, p. 433-440.

Research output: Contribution to journalArticle

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