Purification and characterization of a low molecular weight transforming growth factor from the urine of melanoma patients.

摘要: A low Mr human transforming growth factor (TGF) present in melanoma patients' urine has been purified approximately 200,000-fold to apparent homogeneity. Initial purification of an acid-soluble fraction was achieved by Bio-Gel P-30 gel filtration chromatography 1 M acetic acid. TGF activities were demonstrated the ranges 30,000 and 6,000-10,000. These competed with epidermal (EGF) for binding A431 membrane receptors induced anchorage-independent untransformed fibroblasts. The activity obtained from homogeneity two sequential reverse-phase high performance liquid steps a mu Bondapak C18 column first using linear gradient acetonitrile going 0-60% 120 min then rechromatography over same shallower 20-40% 160 min. isoelectric point patient-derived urinary determined be 6.2, which is distinct that EGF. Amino acid composition analysis (uTGF) revealed it composed at least 42 amino residues minimum estimated 4,545. Compositional further similarities differences between uTGF, EGF TGFs secreted various transformed rodent cell lines.