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J. Biol. Chem., Vol. 265, Issue 25, 14770-14776, 09, 1990
G Elberg, PA Kelly, J Djiane, L Binder and A Gertler
Three monoclonal antibodies (mAbs) (T6, U5, and U6) against prolactin (PRL)
receptors in rat liver were studied in the rat lymphoma lactogen- dependent
(Nb2-11C) and autonomous (Nb2-SP) cell lines. The mAbs had strong affinity
for lactogen receptors (Ka = 12-14 nM-1), similar to that of human growth
hormone (hGH) which is a lactogenic hormone. T6 and hGH competed for the
same binding site, while U5 and U6 interacted with another epitope. The
125I-hGH-receptor complex could be immunoprecipitated by either U5 or U6,
but not by T6. Affinity labeling and immunoblotting revealed that hGH and
U6 bind to a protein of 63-65 kDa. T6, U5, and U6 were mitogenic in Nb2-11C
cells but their respective potencies were 185-, 70-, and 4700-fold lower
than that of hGH. Anti-mouse IgG enhanced the mitogenic effect of all three
mAbs and almost completely abolished the differences between them, although
their mitogenic activity was still 60-120-fold lower than hGH. Des-13- hGH,
a competitive antagonist of hGH which hardly effected the binding of
125I-U5, inhibited the U5-stimulated proliferation of Nb2-11C cells in a
noncompetitive manner, indicating that simultaneous binding of both ligands
fixed the receptor in a nonactive conformation. A Fab fragment of T6 was
not mitogenic, and inhibited the hGH-induced mitogenesis in a competitive
manner, but its mitogenicity could be restored by anti-mouse IgG. We
suggest that the dimerization or oligomerization of the lactogen receptor
in Nb2-11C cells is an obligatory step in the transduction of the mitogenic
signal. It may be induced by binding of the mAb to a site, which can be
either identical or may even be distinct from that which binds the
lactogenic hormone.
Mitogenic and binding properties of monoclonal antibodies to the prolactin receptor in Nb2 rat lymphoma cells. Selective enhancement by anti-mouse IgG
Department of Biochemistry and Human Nutrition, Faculty of Agriculture, Hebrew University of Jerusalem, Rehovot, Israel.
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