Endocrinology, Vol 127, 1167-1175, Copyright © 1990 by Endocrine Society

ARTICLES

Monoclonal and polyclonal antibodies to human progesterone receptor peptide-(533-547) recognize a specific site in unactivated (8S) and activated (4S) progesterone receptor and distinguish between intact and proteolyzed receptors

AM Traish and HH Wotiz
Department of Biochemistry, Boston University School of Medicine, Massachusetts 02118.

We have synthesized three peptides with amino acid sequences corresponding to amino acids 533-547, 597-611, and 765-779 of the human progesterone receptor (hPR). These peptides were conjugated to keyhole limpet hemocyanin and injected into mice and rabbits to develop antibodies to hPR. Antibodies to the undenatured form of PR were elicited only by the peptide with amino acid sequence 533-547. Fusion of SP2/0 myeloma cells with spleen cells from mice immunized with this peptide produced several active clones. Rabbit sera from immunized animals produced one antiserum that reacted with the undenatured form of PR. One monoclonal antibody (PR-AT 4.14) and one antiserum (PR- AT533) raised against peptide-(533-547) were characterized. Binding of these antibodies to the undenatured form of PR was demonstrated by analysis of the antibody-receptor complexes on sucrose density gradients and by immunoprecipitation techniques. Binding of PR to the antibodies was inhibited by excess peptide. The antibodies did not react with estrogen, glucocorticoid, or androgen receptors, but recognized PR from human breast cancer as well as calf, rabbit, mouse, and rat uteri, indicating that this epitope was conserved among these species. Based on sucrose density gradient analysis of PR prepared and labeled in the presence of proteolysis inhibitors and sodium molybdate, the antibodies bound to a site on the intact undenatured PR, but failed to bind to partially degraded steroid-binding form of the receptor, suggesting that the antibody-binding domain is at or near a site sensitive to proteolysis.

This article has been cited by other articles:

				P. S. Quadros and C. K. Wagner Regulation of Progesterone Receptor Expression by Estradiol Is Dependent on Age, Sex and Region in the Rat Brain Endocrinology, June 1, 2008; 149(6): 3054 - 3061. [Abstract] [Full Text] [PDF]

				C. V. V. Helena, M. de Oliveira Poletini, G. L. Sanvitto, S. Hayashi, C. R. Franci, and J. A. Anselmo-Franci Changes in {alpha}-estradiol receptor and progesterone receptor expression in the locus coeruleus and preoptic area throughout the rat estrous cycle J. Endocrinol., February 1, 2006; 188(2): 155 - 165. [Abstract] [Full Text] [PDF]

				T. D. Lund, T. Rovis, W. C. J. Chung, and R. J. Handa Novel Actions of Estrogen Receptor-{beta} on Anxiety-Related Behaviors Endocrinology, February 1, 2005; 146(2): 797 - 807. [Abstract] [Full Text] [PDF]

				A. Guzeloglu, T. R. Bilby, A. Meikle, S. Kamimura, A. Kowalski, F. Michel, L. A. MacLaren, and W. W. Thatcher Pregnancy and Bovine Somatotropin in Nonlactating Dairy Cows: II. Endometrial Gene Expression Related to Maintenance of Pregnancy J Dairy Sci, October 1, 2004; 87(10): 3268 - 3279. [Abstract] [Full Text] [PDF]

				C. K. Wagner, J. Xu, J. L. Pfau, P. S. Quadros, G. J. De Vries, and A. P. Arnold Neonatal Mice Possessing an Sry Transgene Show a Masculinized Pattern of Progesterone Receptor Expression in the Brain Independent of Sex Chromosome Status Endocrinology, March 1, 2004; 145(3): 1046 - 1049. [Abstract] [Full Text] [PDF]

				P. S. Quadros, J. L. Pfau, A. Y. N. Goldstein, G. J. De Vries, and C. K. Wagner Sex Differences in Progesterone Receptor Expression: A Potential Mechanism for Estradiol-Mediated Sexual Differentiation Endocrinology, October 1, 2002; 143(10): 3727 - 3739. [Abstract] [Full Text] [PDF]

				M. Fodor, F. W. van Leeuwen, and D. F. Swaab Differences in Postmortem Stability of Sex Steroid Receptor Immunoreactivity in Rat Brain J. Histochem. Cytochem., May 1, 2002; 50(5): 641 - 650. [Abstract] [Full Text] [PDF]

				M. Jonsson, J. Dejmek, P.-O. Bendahl, and T. Andersson Loss of Wnt-5a Protein Is Associated with Early Relapse in Invasive Ductal Breast Carcinomas Cancer Res., January 1, 2002; 62(2): 409 - 416. [Abstract] [Full Text] [PDF]

				H. Sakamoto, K. Ukena, and K. Tsutsui Effects of Progesterone Synthesized De Novo in the Developing Purkinje Cell on Its Dendritic Growth and Synaptogenesis J. Neurosci., August 15, 2001; 21(16): 6221 - 6232. [Abstract] [Full Text] [PDF]

				U. Ottander, K. Hosokawa, K. Liu, A. Bergh, T. Ny, and J. I. Olofsson A Putative Stimulatory Role of Progesterone Acting via Progesterone Receptors in the Steroidogenic Cells of the Human Corpus Luteum Biol Reprod, March 1, 2000; 62(3): 655 - 663. [Abstract] [Full Text]

				T. E. Spencer, A. Gray, G. A. Johnson, K. M. Taylor, A. Gertler, E. Gootwine, T. L. Ott, and F. W. Bazer Effects of Recombinant Ovine Interferon Tau, Placental Lactogen, and Growth Hormone on the Ovine Uterus Biol Reprod, December 1, 1999; 61(6): 1409 - 1418. [Abstract] [Full Text]

				S. A. Haywood, S. X. Simonian, E. M. van der Beek, R. J. Bicknell, and A. E. Herbison Fluctuating Estrogen and Progesterone Receptor Expression in Brainstem Norepinephrine Neurons through the Rat Estrous Cycle Endocrinology, July 1, 1999; 140(7): 3255 - 3263. [Abstract] [Full Text]

				S. Le Bihan, V. Marsaud, C. Mercier-Bodard, E.-E. Baulieu, S. Mader, J. H. White, and J.-M. Renoir Calcium/Calmodulin Kinase Inhibitors and Immunosuppressant Macrolides Rapamycin and FK506 Inhibit Progestin- and Glucocorticosteroid Receptor-Mediated Transcription in Human Breast Cancer T47D Cells Mol. Endocrinol., July 1, 1998; 12(7): 986 - 1001. [Abstract] [Full Text]