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Horm Metab Res 1992; 24(12): 570-575
DOI: 10.1055/s-2007-1003392
Originals Basic

© Georg Thieme Verlag, Stuttgart · New York

Identification of Müllerian Inhibiting Substance Specific Binding in Human Cell Lines

D. T. MacLaughlin1 , 3 , 6 , R. K. Levin1 , Elizabeth A. Catlin1 , 4 , Lesli A. Taylor1 , F. I. Preffer2 , 5 , Patricia K. Donahoe1
  • 1Pediatric Surgical Research Laboratory, Massachusetts General Hospital, Boston, Massachusetts, U.S.A.
  • 2The Division of Immunopathology, Massachusetts General Hospital, Boston, Massachusetts, U.S.A.
  • 3The Vincent Research Laboratory, Massachusetts General Hospital, Boston, Massachusetts, U.S.A.
  • 4The Departments of Surgery, Pediatrics, Harvard Medical School, Boston, Massachusetts, U.S.A.
  • 5The Department of Pathology, Harvard Medical School, Boston, Massachusetts, U.S.A.
  • 6The Departments of Obstetrics and Gynecology, Harvard Medical School, Boston, Massachusetts, U.S.A.
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Publication History

1991

1992

Publication Date:
14 March 2008 (online)

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Summary

The receptor for Müllerian Inhibiting Substance (MIS), a gonadal glycoprotein hormone, has not been previously identified. Plasma membranes from MlS-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-idiorypic 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.

Key words

Müllerian Inhibiting Substance - Receptor - Anti-idiotypic Antibody - Binder

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