Identification of müllerian inhibiting substance specific binding in human cell lines.
The receptor for Müllerian Inhibiting Substance (MIS), a gonadal glycoprotein hormone, has not been previously identified. Plasma membranes from MIS-sensitive human tumor cell lines (HTB-111, endometrial carcinoma; and A-431, vulvar squamous carcinoma) were detergent extracted and incubated with 125I-labeled MIS anti-idiotypic antibody, or radioiodinated human recombinant MIS (125I rhMIS), with and without unlabeled competitors. 125I anti-idiotypic MIS antibody bound to HTB-111 membrane extracts was displaceable by unlabeled anti-idiotypic antibody, but not by anti-isotypic antibody prior to cross-linking. Specific binding of the anti-idiotypic MIS antibody to endometrial carcinoma cells was verified using fluorescence activated cell analysis and fluoresceinated antibody. Furthermore, unlabeled anti-idiotypic MIS antibody competed for 125I rhMIS binding to A-431 vulvar carcinoma membranes. The labeled anti-idiotypic MIS antibody binding complex could be separated from 32P labeled EGF receptor in the A-431 membranes, indicating that EGF, a natural inhibitor of MIS activity, and MIS itself bind to different receptors. These studies demonstrate a specific, displaceable binder for MIS in the plasmalemmae of two human tumor lines. Purification of this cell surface receptor protein will be greatly aided by using the MIS anti-idiotypic antibody.
MacLaughlin
D T
DT
Pediatric Surgical Research Laboratory, Massachusetts General Hospital, Boston.
Levin
R K
RK
Catlin
E A
EA
Taylor
L A
LA
Preffer
F I
FI
Donahoe
P K
PK
D054304:Anti-Mullerian Hormone; D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D016889:Endometrial Neoplasms / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D006023:Glycoproteins; D006131:Growth Inhibitors / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D007457:Iodine Radioisotopes; D011994:Recombinant Proteins / Q000378:metabolism; D013735:Testicular Hormones / Q000276:immunology / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014846:Vulvar Neoplasms / Q000378:metabolism*
D000906:Antibodies; D006023:Glycoproteins; D006131:Growth Inhibitors; D007130:Immunoglobulin Idiotypes; D007457:Iodine Radioisotopes; D011994:Recombinant Proteins; D013735:Testicular Hormones; D054304:Anti-Mullerian Hormone