Amino-terminal deletion of heparin-binding epidermal growth factor-like growth factor4-127 stimulates cell proliferation but lacks insulin-like activity.

UNLABELLED: Heparin-binding epidermal growth factor-like growth factor (HB-EGF) Northern analysis demonstrated a novel 0.8-kb liver-specific HB-EGF transcript in addition to the endogenous 2.5-kb HB-EGF full-length transcript present in kidney, lung and liver tissues. Reverse transcriptase-polymerase chain reaction screening of liver RNA suggests that the 0.8-kb HB-EGF transcript lacks at least a portion of the amino-terminal EGF-like domain. In light of these data, we have constructed a human HB-EGF cDNA (HB-EGF(DeltaN)) which lacks 373 bp, encoding the majority of the extracellular EGF-like domain, while maintaining the ectodomain 'shedding' site, transmembrane and cytoplasmic domains.
OBJECTIVE: The goal of this study is to characterize the ability of HB-EGF(DeltaN) to (i) stimulate cell proliferation and (ii) determine whether down-regulation of insulin-like growth factor-binding protein (IGFBP)-3 and -4 mRNA is regulated by soluble, mature HB-EGF or HB-EGF C.
MATERIALS AND METHODS: HB-EGF(DeltaN) encodes nucleotides +1-10 of exon 1 linked to nucleotides 383-627 of the carboxy-terminal portion of exon 3 through exon 5.
RESULTS: Expression of HB-EGF(DeltaN) in mouse fibroblasts (MLC) resulted in 6.5- and 8-kDa HB-EGF immunoreactive proteins, stimulated tyrosine phosphorylation of p42 kDa and cell proliferation in MLC, but lacked the ability to bind EGF receptors. Finally, HB-EGF(DeltaN) failed to down-regulate IGFBP-3 and -4 mRNA when expressed in normal rat kidney cells.
CONCLUSIONS: These findings demonstrate that amino-terminally truncated, membrane-bound form of HB-EGF stimulates cell proliferation but lacks insulin-like signalling, suggesting that insulin-like signalling is mediated by soluble, mature HB-EGF binding to EGF receptors.