Murine Epidermal Growth Factor: Structure and Function

Antony W. Burgess, Christopher J. Lloyd, Sandra Smith, Edouard Stanley, Francesca Walker, Louis Fabri, Richard J. Simpson, Edouard C. Nice

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Murine epidermal growth factor (EGF), a 53 amino acid protein, has been modified by enzymic digestion, site-specific chemical reactions, and recombinant DNA technology. After trypsin digestion the EGF derivatives EGF1-48(called EGF-T) and EGF1-45(called EGF-T2) were separated from the residual EGF and the C-terminal pentapeptide by reversed-phase high-performance liquid chromatography. EGF-T competes for binding to EGF receptors with the same efficiency as EGF. the EGF-T2derivative had no detectable receptor binding activity even at 100 nM. the in vitro mitogenic potencies of EGF and EGF-T for Balb/c 3T3 cells were indistinguishable. Treatment of EGF-T with carboxypeptidase Y yielded two derivatives, EGF-T-(des-Arg48) and EGF-T-des(Leu47-Arg48). There was only a 3-7-fold diminution in the binding efficiency and mitogenic potency for EGF-T-(des-Arg48). However, there was more than a 100-fold decrease in the binding efficiency and mitogenic activity of EGF-T-des(Leu47-Arg48). These results indicated that Leu47is intimately involved in the formation of the ligand-receptor complex. Studies with a number of proteases indicated that the C-terminus of EGF was susceptible to enzymic digestion; however, the N-terminus appears to be folded into a conformation which prevents access to proteolytic digestion. Consequently, the N-terminus was modified by preparing an analogue with recombinant DNA technology. Oligonucleotides corresponding to EGF(3_48).Met3.Lys21residues were ligated in frame to a β-galactosidase expression vector. the β-Gal-EGF fusion protein was cleaved with cyanogen bromide and EGF(4_48).Lys21purified. This derivative was equipotent with EGF in the mitogenesis assay and bound to the EGF receptor with the same affinities as EGF. Disruption of the central antiparallel β-sheet structure of EGF at Met21by treatment of EGF with cyanogen bromide reduced both the binding efficiency and the mitogenic activity of EGF more than 100-fold.

Original languageEnglish
Pages (from-to)4977-4985
Number of pages9
JournalBiochemistry
Volume27
Issue number14
DOIs
Publication statusPublished - 1 Jul 1988
Externally publishedYes

Cite this

Burgess, A. W., Lloyd, C. J., Smith, S., Stanley, E., Walker, F., Fabri, L., ... Nice, E. C. (1988). Murine Epidermal Growth Factor: Structure and Function. Biochemistry, 27(14), 4977-4985. https://doi.org/10.1021/bi00414a005
Burgess, Antony W. ; Lloyd, Christopher J. ; Smith, Sandra ; Stanley, Edouard ; Walker, Francesca ; Fabri, Louis ; Simpson, Richard J. ; Nice, Edouard C. / Murine Epidermal Growth Factor : Structure and Function. In: Biochemistry. 1988 ; Vol. 27, No. 14. pp. 4977-4985.
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Burgess, AW, Lloyd, CJ, Smith, S, Stanley, E, Walker, F, Fabri, L, Simpson, RJ & Nice, EC 1988, 'Murine Epidermal Growth Factor: Structure and Function' Biochemistry, vol. 27, no. 14, pp. 4977-4985. https://doi.org/10.1021/bi00414a005

Murine Epidermal Growth Factor : Structure and Function. / Burgess, Antony W.; Lloyd, Christopher J.; Smith, Sandra; Stanley, Edouard; Walker, Francesca; Fabri, Louis; Simpson, Richard J.; Nice, Edouard C.

In: Biochemistry, Vol. 27, No. 14, 01.07.1988, p. 4977-4985.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Burgess, Antony W.

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Burgess AW, Lloyd CJ, Smith S, Stanley E, Walker F, Fabri L et al. Murine Epidermal Growth Factor: Structure and Function. Biochemistry. 1988 Jul 1;27(14):4977-4985. https://doi.org/10.1021/bi00414a005