A 40-kDa epidermal growth factor/transforming growth factor α-binding domain produced by limited proteolysis of the extracellular domain of the epidermal growth factor receptor

D. Kohda, M. Odaka, I. Lax, H. Kawasaki, K. Suzuki, A. Ullrich, J. Schlessinger, F. Inagaki

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Elucidation of the three-dimensional structure of the complex of the epidermal growth factor (EGF) and its receptor is essential for understanding the molecular mechanisms of the EGF-receptor interaction and EGF-induced receptor-receptor interaction. NMR is useful to investigate interactions in solution between macromolecules at atomic resolution, but has a limitation in molecular masses of target proteins: less than 300 residues. We have prepared a fragment with apparent molecular mass of 40 kDa in SDS gels from the soluble extracellular domain of the EGF receptor (sEGFR, 619 residues) by sequential limited proteolysis with proteinase K and bromelain. This fragment is a monomeric structural domain consisting of 202 amino acid residues (Cys302-Arg503) and 18-kDa sugar chains, and binds EGF and transforming growth factor-α (TGFα). This 40-kDa domain has a dissociation constant of about 1 μM for human TGFα, which is similar to that of the parental sEGFR. sEGFR oligomerizes in response to EGF and TGFα, while the 40-kDa domain does not, suggesting that the sequences other than this domain is required for receptor oligomerization. The 40-kDa ligand-binding domain described in this report is suitable for analysis by various physico-chemical approaches such as NMR.

Original languageEnglish
Pages (from-to)1976-1981
Number of pages6
JournalJournal of Biological Chemistry
Volume268
Issue number3
Publication statusPublished - Jan 1 1993
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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