Inhibin binding sites and proteins in pituitary, gonadal, adrenal and bone cells.
Activin signals via complexes of type I (50-55 kDa) and II (70-75 kDa) activin receptors, but the mechanism of inhibin action is unclear. Proposed models range from an anti-activin action at the type II activin receptor to independent actions involving putative inhibin receptors. Two membrane-embedded proteoglycans, betaglycan and p120, have recently been implicated in inhibin binding, but neither appears to be a signalling receptor. The present studies on primary cultures of rat pituitary and adrenal cells, and several murine and human cell lines were undertaken to characterise inhibin binding to its physiological targets. High affinity binding of inhibin to the primary cultures and several of the cell lines, like that previously described for ovine pituitary cells, was saturable and reversible. Scatchard analysis revealed two classes of binding sites (K(d) of 40-400 and 500-5000 pM, respectively). Affinity labelling identified [125I]inhibin binding proteins with apparent molecular weights of 41, 74, 114 and >170 kDa in all cell types that displayed high affinity, high capacity binding of inhibin. Additional labelling of a 124 kDa species was evident in gonadal TM3 and TM4 cell lines. In several cases, activin (> or =20 nM) competed poorly or not at all for binding to these proteins. The 74, 114 and >170 kDa inhibin binding proteins in TM3 and TM4 cells were immunoprecipitated by an anti-betaglycan antiserum. These three proteins correspond in size to the activin receptor type II and the core protein and glycosylated forms of betaglycan, respectively, that have been proposed to mediate anti-activin actions of inhibin, but the identity of the 74 kDa species is yet to be confirmed. Studies of [125I]inhibin binding kinetics and competition for affinity labelling of individual binding proteins in several cell lines suggest these three species and the 41 and 124 kDa proteins form a high affinity inhibin binding complex. In summary, common patterns of inhibin binding and affinity labelling were observed in inhibin target cells. Novel inhibin binding proteins of around 41 and 124 kDa were implicated in the high affinity binding of inhibin to cells from several sources.
Farnworth
P G
PG
Prince Henry's Institute of Medical Research, P.O. Box 5152, 3168, Victoria, Clayton, Australia. paul.farnworth@med.monash.edu.au
Harrison
C A
CA
Leembruggen
P
P
Chan
K L
KL
Stanton
P G
PG
Ooi
G T
GT
Rahman
N A
NA
Huhtaniemi
I T
IT
Findlay
J K
JK
Robertson
D M
DM
D029404:Activin Receptors; D000311:Adrenal Glands / Q000166:cytology / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001842:Bone and Bones / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D006066:Gonads / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007265:Inhibins / Q000378:metabolism*; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism; D011485:Protein Binding; D018000:Receptors, Peptide / Q000378:metabolism*
D018000:Receptors, Peptide; D007265:Inhibins; D029404:Activin Receptors