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Isolation and characterization of epidermal growth factor from human milk
Author(s) -
Petrides Petro E.,
Hosang Markus,
Shooter Eric,
Esch Frederic S.,
Böhlen Peter
Publication year - 1985
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(85)81220-5
Subject(s) - isolation (microbiology) , epidermal growth factor , characterization (materials science) , chemistry , biology , biochemistry , microbiology and biotechnology , nanotechnology , materials science , receptor
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 NH 2 ‐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 NH 2 ‐terminof urogastrone from human urine. The purified polypeptide competes with mEGF for the placental membrane receptor with a k i , 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 , 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.