THE RELATIVE IMPORTANCE OF THE OLIGOSACCHARIDE UNITS IN HUMAN CHORIONIC-GONADOTROPIN (CG) FOR LH CG RECEPTOR ACTIVATION IN RAT LEYDIG-CELLSAND MOUSE LEYDIG TUMOR-CELLS/
Hj. Vanloenen et al., THE RELATIVE IMPORTANCE OF THE OLIGOSACCHARIDE UNITS IN HUMAN CHORIONIC-GONADOTROPIN (CG) FOR LH CG RECEPTOR ACTIVATION IN RAT LEYDIG-CELLSAND MOUSE LEYDIG TUMOR-CELLS/, Journal of Endocrinology, 147(2), 1995, pp. 367-375
The biological properties of deglycosylated human chorionic gonadotrop
in (DhCG), obtained by hydrogen fluoride treatment (HF-DhCG) of intact
hCG or by oligonucleotide-directed mutagenesis (CHO-DhCG), and that o
f their fully glycosylated counterparts, were tested in term of cAMP a
nd steroid production in rat Leydig cells and in mouse Leydig tumor ce
lls (MA-10 cells). In both cell types, HF-DhCG and CHO-DhCG possessed
comparable biological activities. The maximum for DhCG-induced cAMP pr
oduction was approximately 12% of that of intact hCG when tested in ra
t Leydig cells, and only 2% when tested in MA-10 cells. DhCG possessed
significant steroidogenic activity in both cell types. In MA-10 cells
the maximum for DhCG-induced steroidogenesis was 30-50% of that of in
tact hCG, while in rat Leydig cells DhCG and hCG induced similar stero
idogenic maxima. Based on its ED(50), DhCG possessed 10-17% of the ste
roidogenic potency of intact hCG in rat Leydig cells, while in MA-10 c
ells DhCG was only a-fold less potent than hCG. When accurate hormone-
receptor binding data are absent, the intrinsic receptor-stimulating a
ctivity of a ligand can still be estimated at full receptor occupancy,
provided that over the whole dose range the biological response is pr
oportional to receptor stimulation. The present data show that in tran
sfected MA-10(P(+)29) cells which over-express rat phosphodiesterase,
the hormone-induced stimulation of cAMP and steroid production is dire
ctly coupled to receptor activation up to maximal occupation of the LH
/CG receptor. The intrinsic receptor-stimulating activity of DhCG, mea
sured in MA-10(P(+)29) cells in terms of DhCG-induced steroidogenesis,
appeared to be 7- to 10-fold lower than that of intact hCG. As it is
known from the Literature that DhCG possesses 2- to 3-fold higher affi
nity to the LH/CG receptor than intact hCG, this increased binding aff
inity of DhCG may partly compensate for the 7- to 10-fold reduction in
the intrinsic receptor-stimulating activity, resulting in only a 2-fo
ld reduction in steroidogenic potency of DhCG. In terms of adenylyl cy
clase stimulation in MA-10(P(+)29) cells, DhCG possessed approximately
50-fold lower receptor-stimulating activity than intact hCG, being si
milar to that observed in wild-type MA-10 cells. This study clearly sh
ows that the oligosaccharide units in hCG are not essential for LH/CG
receptor activation, and that the relative receptor-stimulating activi
ty of DhCG to that of. hCG is highly dependent on whether cAMP or ster
oid production is measured as an index for bioactivity, and whether bi
oactivity is tested in rat Leydig cells or MA-10 cells.