Chemical synthesis and biological activity of the EGF-like domain of heparin-binding epidermal growth factor-like growth factor (HB-EGF).
Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a recently discovered member of the epidermal growth factor (EGF) family. This novel growth factor possesses the EGF-like domain in the carboxyl portion. In order to evaluate the biological function of the EGF-like domain in HB-EGF, human HB-EGF(44-86) corresponding to the EGF-like domain was synthesized by the solid-phase procedure using the Fmoc strategy. It was confirmed by amino acid microsequencing of cysteine-containing fragments derived from thermolytic digestion that the pattern of three disulfide bond pairings in synthetic HB-EGF(44-86) was consistent with that of EGF and transforming growth factor-alpha (TGF-alpha). The homogeneity of the synthetic peptide was confirmed by analytical RP-HPLC, amino acid analysis and fast atom bombardment mass spectrometer (FAB-MS). Compared with h-EGF, the EGF-like domain of human HB-EGF showed a comparable mitogenic activity in the proliferation of NIH/3T3 fibroblast cells. These results suggest that the EGF-like domain of human HB-EGF may play an important role in mitogenic activity.
Shin
S Y
SY
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
Takenouchi
T
T
Yokoyama
T
T
Ohtaki
T
T
Munekata
E
E
D016475:3T3 Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D003124:Colorimetry; D004815:Epidermal Growth Factor / Q000138:chemical synthesis* / Q000494:pharmacology*; D005449:Fluorenes / Q000737:chemistry; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013778:Tetrazolium Salts; D013936:Thymidine / Q000378:metabolism; D014269:Trifluoroacetic Acid / Q000737:chemistry; D014316:Tritium
D000596:Amino Acids; D005449:Fluorenes; C587883:HBEGF protein, human; C587884:Hbegf protein, mouse; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D010446:Peptide Fragments; D013778:Tetrazolium Salts; D014316:Tritium; D004815:Epidermal Growth Factor; D014269:Trifluoroacetic Acid; D013936:Thymidine