The interaction of an epidermal growth factor transforming growth factor alpha tail chimera with the human epidermal growth factor receptor reveals unexpected complexities

被引:13
作者
Puddicombe, SM [1 ]
Wood, L [1 ]
Chamberlin, SG [1 ]
Davies, DE [1 ]
机构
[1] SOUTHAMPTON GEN HOSP, CANC RES CAMPAIGN MED ONCOL UNIT, SOUTHAMPTON SO16 6YD, HANTS, ENGLAND
基金
英国惠康基金;
关键词
D O I
10.1074/jbc.271.48.30392
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
It has been assumed that substitution of homologous regions of transforming growth factor alpha (TGF-alpha) into epidermal growth factor (EGF) can be used to probe ligand-receptor recognition without detrimental effects on ligand characteristics for the human EGF receptor (EGFR), We show that a chimera of murine (m) EGF in which the carboxyl-terminal tail is substituted for that of TGF-alpha (mEGF/TGF-alpha(44-50)) results in complex features that belie this initial simplistic assumption, Comparison of EGF and mEGF/TGF-alpha(44-50) in equilibrium binding assays showed that although the relative binding affinity of the chimera was reduced 80-200-fold, it was more potent than EGF in mitogenesis assays using NR6/HER cells, This superagonist activity could not be attributed to differences in ligand processing or to binding to other members of the c-erbB family. It appeared to be due, in part, to choice of an EGFR overexpressing target cell where high receptor number compensated for the low affinity of the ligand; it also appeared to be related to the ability of the chimera to activate the EGFR tyrosine kinase, Thus, when EGFR autophosphorylation was measured, mEGF/TGF-alpha(44-50) was more potent than EGF, despite its low affinity, When tested using chicken embryo fibroblasts, substitution of the TGF-alpha carboxyl-terminal tail into mEGF failed to enhance its binding affinity for chicken EGFRs; however, the chimera was intermediate in potency between TGF-alpha and mEGF in mitogenesis assays, Our results suggest a contextual requirement for EGFR recognition which is ligand-specific, Further, the unpredictable responses to chimeric ligands underline the complex nature of the processes of ligand recognition, receptor activation, and the ensuing cellular response.
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页码:30392 / 30397
页数:6
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