Endocrinology, Vol 131, 291-296, Copyright © 1992 by Endocrine Society

ARTICLES

Mullerian duct regression and antiproliferative bioactivities of mullerian inhibiting substance reside in its carboxy-terminal domain

DT MacLaughlin, PL Hudson, AL Graciano, MK Kenneally, RC Ragin, TF Manganaro and PK Donahoe
Pediatric Surgical Research Laboratories, Massachusetts General Hospital, Boston 02114.

A 25-kilodalton dimeric carboxy-terminal fragment of the recombinant human Mullerian inhibiting substance protein (rhMIS) was produced by proteolytic cleavage with plasmin and purified by size-exclusion chromatography. The identity of the isolated dimer as the carboxy- terminal fragment was confirmed by gel electrophoresis and Western analysis. As was true of every sample of the holo molecule, all preparations of the carboxy-terminal domain of rhMIS (n = 10), when added in the 0.5-5.0 micrograms/ml range, exhibited a dose-dependent partial to complete regression of the 14.5-day fetal rat Mullerian duct in an organ culture assay. The carboxy-terminal dimer also inhibited, in a dose-dependent manner, the growth of A431 cells in monolayer cultures. Daily addition of 5, 10, or 20 micrograms carboxy-terminus for 3 days resulted in 0%, 25%, and 100% inhibition of cell proliferation, respectively. Similar and higher doses of holo rhMIS had no or inconsistent antiproliferative activity (0-34% inhibition), even though the preparations caused Mullerian duct regression. All amino- terminal fragments prepared using this separation protocol were found to be inactive in these assays. These findings suggest that the bioactivity of rhMIS as a regressor of fetal Mullerian ducts and an inhibitor of A431 cell growth resides in its carboxy-terminal domain. These results indicate that the urogenital ridge tissue, but not A431 cells in culture, may be capable of cleaving intact MIS to a biologically active conformation.

This article has been cited by other articles:

				J. Teixeira, S. Maheswaran, and P. K. Donahoe Mullerian Inhibiting Substance: An Instructive Developmental Hormone with Diagnostic and Possible Therapeutic Applications Endocr. Rev., October 1, 2001; 22(5): 657 - 674. [Abstract] [Full Text] [PDF]

				P. T. Masiakos, D. T. MacLaughlin, S. Maheswaran, J. Teixeira, A. F. Fuller Jr., P. C. Shah, D. J. Kehas, M. K. Kenneally, D. M. Dombkowski, T. U. Ha, et al. Human Ovarian Cancer, Cell Lines, and Primary Ascites Cells Express the Human Mullerian Inhibiting Substance (MIS) Type II Receptor, Bind, and Are Responsive to MIS Clin. Cancer Res., November 1, 1999; 5(11): 3488 - 3499. [Abstract] [Full Text] [PDF]

				J. Teixeira, E. Fynn-Thompson, A. H. Payne, and P. K. Donahoe Mullerian-Inhibiting Substance Regulates Androgen Synthesis at the Transcriptional Level Endocrinology, October 1, 1999; 140(10): 4732 - 4738. [Abstract] [Full Text]

				J. M. Hutson, S. Hasthorpe, and C. F. Heyns Anatomical and Functional Aspects of Testicular Descent and Cryptorchidism Endocr. Rev., April 1, 1997; 18(2): 259 - 280. [Abstract] [Full Text]

				E. A. Catlin, V. C. Tonnu, R. G. Ebb, B. A. Pacheco, T. F. Manganaro, R. M. Ezzell, P. K. Donahoe, and J. Teixeira Mullerian Inhibiting Substance Inhibits Branching Morphogenesis and Induces Apoptosis in Fetal Rat Lung Endocrinology, February 1, 1997; 138(2): 790 - 796. [Abstract] [Full Text] [PDF]