Purification and properties of epithelial growth inhibitor (EGI) from human platelets: its separation from type beta transforming growth factor (TGF-beta).
We previously reported that sera from various kinds of animals contain a protein(s) capable of inhibiting the growth of the non-malignant epithelial cell line derived from Buffalo rat liver (BRL). In the present study, a similar epithelial cell-specific growth inhibitor (EGI) was purified to homogeneity from an acid-ethanol extract of human platelets. During purification, EGI was separated from the major component of type beta transforming growth factor (TGF-beta), which can stimulate the colony formation of the non-malignant fibroblastic cell line derived from rat kidney (NRK) in soft agar in the presence of epidermal growth factor (EGF). The purified EGI had an Mr of 27,000, and was composed of two subunits identical in Mr. It significantly inhibited the growth in monolayer cultures of three non-malignant epithelial cell lines, BRL, MDCK (from Madin-Darby canine kidney) and BSC-1 (from African green monkey kidney), at doses lower than 40 pg/ml in medium containing 10% fetal calf serum. Its inhibitory activity was stable against heating at 90 degrees C for 3 min, but not against treatment with 50 mM dithiothreitol. In addition, TGF-beta was also partially purified from the same extract. The purified TGF-beta did not show any inhibitory activity toward the growth of BRL, MDCK, BSC-1, or NRK.
Huang
W
W
Kimura
T
T
Mashima
K
K
Miyazaki
K
K
Masaki
H
H
Yamashita
J
J
Horio
T
T
D001792:Blood Platelets / Q000032:analysis*; D002460:Cell Line; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006131:Growth Inhibitors / Q000032:analysis / Q000097:blood / Q000302:isolation & purification*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis / Q000097:blood / Q000302:isolation & purification*; D010455:Peptides / Q000032:analysis / Q000097:blood / Q000302:isolation & purification*; D015291:Transforming Growth Factors
D006131:Growth Inhibitors; D008565:Membrane Proteins; D010455:Peptides; C031717:growth inhibitory proteins; D015291:Transforming Growth Factors