Chemical synthesis and biological activities of the EGF-like domain of mouse schwannoma-derived growth factor (SDGF).
Schwannoma-derived growth factor (SDGF), an epidermal growth factor (EGF) family peptide recently discovered, has an EGF-like domain in the carboxyl terminal portion. In this study, we synthesized mSDGF(38-80) corresponding to the EGF-like domain of mouse SDGF by means of stepwise solid-phase method using Fmoc chemistry in order to evaluate the biological function of the EGF-like domain in SDGF. The linear peptide of mSDGF(38-80) was folded by direct oxidation with reduced and oxidized glutathione to form intramolecular disulfide bridges in synthetic peptide. On the biological activity, we examined mitogenic activity induced by mSDGF(38-80) in NIH/3T3 fibroblast cells and interaction with EGF receptor in A431 cells. In the results, mSDGF(38-80) was confirmed to form three disulfide linkages that were similar in pattern to EGF by amino acid and sequence analysis of fragments obtained after thermolytic digestion. However, mSDGF(38-80) possessed weak mitogenic activity in NIH/3T3 cells and weak binding affinity for the EGF receptor in A431 cells compared with those of human EGF. These results suggest that the EGF-like domain of SDGF may have little effect upon mitogenic activity and the EGF receptor binding of SDGF.
Takenouchi
T
T
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
Kadosaka
M
M
Shin
S Y
SY
Munekata
E
E
D016475:3T3 Cells; D000595:Amino Acid Sequence; D066258:Amphiregulin; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000737:chemistry; D066255:EGF Family of Proteins; D004815:Epidermal Growth Factor / Q000737:chemistry*; D066246:ErbB Receptors / Q000378:metabolism; D006023:Glycoproteins; D006133:Growth Substances / Q000737:chemistry*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D051379:Mice; D008969:Molecular Sequence Data; D009442:Neurilemmoma / Q000737:chemistry*; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology*; D017510:Protein Folding; D017421:Sequence Analysis; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013329:Structure-Activity Relationship; D013820:Thermolysin / Q000378:metabolism
C587848:AREG protein, human; D066258:Amphiregulin; C587849:Areg protein, mouse; D004220:Disulfides; D066255:EGF Family of Proteins; D006023:Glycoproteins; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D010446:Peptide Fragments; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D013820:Thermolysin