Structure and expression of a membrane component of the inhibin receptor system.
The purification and cloning of a membrane-anchored proteoglycan with affinity for inhibin A are described. Bovine pituitary membranes were isolated, and membrane-anchored proteins were solubilized and used as an enriched source of inhibin binding protein. The extract was passed over an inhibin A affinity column, and a protein, designated p120, was identified as an inhibin-binding moiety. A partial amino acid sequence was determined for the protein, which matched two human complementary DNAs (cDNAs) in the database. The full-length cDNA predicts a 1336-amino acid glycoprotein. Full-length p120-encoding cDNAs were isolated from human testis RNA and cloned into expression vectors. Two p120 messenger RNA transcripts of 4.6 kb and 2 kb are detected in rat pituitary by RNA blot analysis. Similar analysis of rat testis RNA revealed transcripts of identical molecular mass, albeit at lower abundance. To determine the cellular localization of p120 in pituitary and testis, an antibody directed against the predicted extracellular domain of the protein was generated and used in an immunohistochemical analysis of thin tissue sections. p120 immunostaining is coincident with FSHbeta immunopositive gonadotrope cells in rat pituitary. p120 staining is intense in the testicular Leydig cells, which bind iodinated inhibin but not iodinated activin. In summary, an inhibin-binding protein has been isolated that is produced in tissues that are targets of inhibin action.
Chong
H
H
Department of Neurobiology and Physiology, Northwestern University, Evanston, Illinois 60208-2850, USA.
Pangas
S A
SA
Bernard
D J
DJ
Wang
E
E
Gitch
J
J
Chen
W
W
Draper
L B
LB
Cox
E T
ET
Woodruff
T K
TK
D029404:Activin Receptors; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008566:Membranes / Q000378:metabolism; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D010902:Pituitary Gland / Q000378:metabolism*; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D051381:Rats; D018000:Receptors, Peptide / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D011509:Proteoglycans; D018000:Receptors, Peptide; D029404:Activin Receptors