Cytotoxicity and specificity of directed toxins composed of diphtheria toxin and the EGF-like domain of heregulin beta1.

Abstract

As a step in the design of directed toxins, aimed at cells that overexpress HER receptors, particularly breast carcinoma cells, we studied the properties of a chimera of diphtheria toxin (DT) and heregulin beta1. The EGF-like growth hormone heregulin is a ligand for the HER3 and HER4 receptors and their heterodimers with HER2. The 60-residue EGF-like domain (hrg) of heregulin elicits a biological response and binds to these receptors primarily through its N terminus. We tested a fusion protein in which hrg replaces the C-terminal receptor-binding domain of DT (DT(389)hrg) and an alternative design in which this domain is fused to the N terminus of DT(389). Of those two constructs, the N-terminal fusion was not active as a directed toxin but elicited a growth response. The C-terminal fusion of hrg to DT(389) yielded a functional toxin and showed cell line specific cytotoxicity that is consistent with heregulin specificity. The binding of hrg to its cognate receptor is not impaired as shown by receptor activation, direct binding, and competition with free hrg. Cytotoxicity is dependent on high-affinity binding of DT(389)hrg to HER3 and HER4 receptors and is not mediated by HER2 overexpression alone. For those cell lines exhibiting high-affinity binding sites, the level of cytotoxicity correlates with the rate of internalization. Thus DT(389)hrg chimeras offer a possible avenue toward directed toxins against cells that overexpress HER receptors.

• Authors: R Landgraf, M Pegram, D J Slamon, D Eisenberg
• Journal: Biochemistry (Biochemistry) Vol. 37 Issue 9 Pg. 3220-8 (Mar 03 1998) ISSN: 0006-2960 [Print] United States
• PMID: 9485477 (Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Carrier Proteins
• Diphtheria Toxin
• Glycoproteins
• Neuregulin-1
• Proto-Oncogene Proteins
• Recombinant Fusion Proteins
• heregulin beta1
• ERBB4 protein, human
• ErbB Receptors
• Receptor, ErbB-2
• Receptor, ErbB-3
• Receptor, ErbB-4

Topics

• Binding Sites
• Carrier Proteins (genetics, metabolism)
• Cell Line
• Cell Survival (drug effects)
• Crystallography, X-Ray
• Diphtheria Toxin (genetics)
• Drug Design
• ErbB Receptors (metabolism)
• Glycoproteins (genetics, metabolism)
• Humans
• Models, Molecular
• Neuregulin-1
• Proto-Oncogene Proteins (metabolism)
• Receptor, ErbB-2 (metabolism)
• Receptor, ErbB-3
• Receptor, ErbB-4
• Recombinant Fusion Proteins (chemical synthesis, metabolism, toxicity)