Isolation and characterization of epidermal growth factor from human milk.
Epidermal growth factor (EGF) has been purified from human milk. The purification was monitored with a human placental membrane radioreceptor assay using murine salivary epidermal growth factor I (mEGF I) as a competitive ligand and was achieved exclusively by the use of reverse-phase liquid chromatography (RPLC). The sequential use of preparative, semipreparative and analytical RPLC on an octylsilica support with solvent systems of different solute selectivity such as pyridine formate, triethylammonnium phosphate or perfluorocarbonic acids in the presence of n-propanol or acetonitrile allowed purification to homogeneity with 5 consecutive runs. The molecular mass, amino acid composition and NH2-terminal sequence of human EGF were determined. Gas-phase microsequencing of residues 1-17 revealed the following sequence: Asn-Ser-Asp-Ser-Glu-X-Pro-Leu-Ser-His-Asp-Gly-Tyr-X-Leu-X-Asp which is identical with the NH2-terminof urogastrone from human urine. The purified polypeptide competes with mEGF for the placental membrane receptor with a ki of 1 ng. Furthermore, it stimulates the anchorage-dependent as well as -independent proliferation of human and rat indicator cells with half-maximal stimulation at 1 and 2.5 ng/ml, respectively. Although human epidermal growth factor has been unequivocally identified in human milk and -for the first time-shown to be identical with urogastrone from human urine, the high-resolution techniques employed have also revealed the presence of EGF-related molecules which await further characterization. It is possible that EGF and the EGF-related growth factors possess important regulatory functions in normal growth of the human breast during pregnancy and lactation as well as in abnormal growth during mammary tumor formation and progression.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001667:Binding, Competitive; D001681:Biological Assay; D004815:Epidermal Growth Factor / Q000302:isolation & purification* / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D008895:Milk, Human / Q000032:analysis*; D010455:Peptides / Q000032:analysis; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D011869:Radioligand Assay; D012463:Saliva / Q000032:analysis; D015291:Transforming Growth Factors
D000596:Amino Acids; D010455:Peptides; D004815:Epidermal Growth Factor; D015291:Transforming Growth Factors