Identification of diphtheria toxin R domain mutants with enhanced inhibitory activity against HB-EGF.

Abstract:

:Heparin-binding epidermal growth factor-like growth factor (HB-EGF), a ligand of EGF receptor, is involved in the growth and malignant progression of cancers. Cross-reacting material 197, CRM197, a non-toxic mutant of diphtheria toxin (DT), specifically binds to the EGF-like domain of HB-EGF and inhibits its mitogenic activity, thus CRM197 is currently under evaluation in clinical trials for cancer therapy. To develop more potent DT mutants than CRM197, we screened various mutant proteins of R domain of DT, the binding site for HB-EGF. A variety of R-domain mutant proteins fused with maltose-binding protein were produced and their inhibitory activity was evaluated in vitro. We found four R domain mutants that showed much higher inhibitory activity against HB-EGF than wild-type (WT) R domain. These R domain mutants suppressed HB-EGF-dependent cell proliferation more effectively than WT R domain. Surface plasmon resonance revealed their higher affinity to HB-EGF than WT R domain. CRM197(R460H) carrying the newly identified mutation showed increased cell proliferation inhibitory activity and affinity to HB-EGF. These results suggest that CRM197(R460H) or other recombinant proteins carrying newly identified mutation(s) in the R domain are potential therapeutics targeting HB-EGF.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Suzuki K,Mizushima H,Abe H,Iwamoto R,Nakamura H,Mekada E

doi

10.1093/jb/mvu079

subject

Has Abstract

pub_date

2015-05-01 00:00:00

pages

331-43

issue

5

eissn

0021-924X

issn

1756-2651

pii

mvu079

journal_volume

157

pub_type

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