Synthesis and biological activity of N-terminal-truncated derivatives of human epidermal growth factor (h-EGF).
To investigate the contribution of the N-terminal sequence of h-EGF to its biological activity and the formation of three intramolecular disulfide bonds by oxidative refolding via air oxidation, five derivatives of h-EGF with a single N-terminal amino acid deletion were synthesized by solid-phase synthesis. The homogeneity of the synthetic peptides was confirmed by analytical reversed-phase HPLC, amino acid analysis, and FAB-MS. The pairing of the three disulfide bridges in synthetic peptides was determined by thermolytic digestion. All N-truncated derivatives of h-FGF formed the correct intramolecular three disulfide linkages during oxidative refolding and had equipotent activity in both EGF receptor binding on A-431 epidermoid carcinoma cells and mitogenesis on NIH-3T3 fibroblast cells, compared with authentic h-EGF. The results suggested that the five residues from N-terminal sequence of h-EGF have no effect on the formation of the correct disulfide linkages in h-EGF and do not exert a significant influence on its biological activity.
Shin
S Y
SY
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
Shimizu
M
M
Ohtaki
T
T
Munekata
E
E
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002294:Carcinoma, Squamous Cell; D002455:Cell Division / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine; D004305:Dose-Response Relationship, Drug; D004815:Epidermal Growth Factor / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D066246:ErbB Receptors / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D011487:Protein Conformation; D017510:Protein Folding; D014407:Tumor Cells, Cultured
D007202:Indicators and Reagents; D010446:Peptide Fragments; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D003545:Cysteine