Our discovery of p59 (FKBP59) was made possible by development of cell line KN382/EC1, a mouse hybridoma secreting an IgG, directed against the nontransformed progestin receptor. It is present in rabbit uterus, kidney, and liver, calf uterus, Cos cells, human breast cancer cells, human lymphoma cells, and monkey uterus. Identification of p59 as FKBP59, an immunophilin, was accomplished by collaboration between our laboratory and that of Dr. Stewart Schreiber (see letter of collaboration). P59 (FKBP59) is an intracellular receptor for FK506 and rapamycin. We know that P59 is ubiquitous, associates with heat shock proteins, is in excess of, and a component of, nontransformed steroid receptors. It may be a heat shock protein. We believe p59 (FKBP59) is an important intermediate between the immunosuppressants and steroids, involved in protein trafficking and signal transduction. To prove or disprove this, we shall, 1. Determine intracellular targets for p59 (FKBP59). 2. Determine the binding and enzymatic characteristics of p59 (FKBP59). 3. Determine the endocrine (biological control) of p59 (FKBP59). We shall accomplish the first of our goals by means of KN382/EC1, FK506, and p59 affinity chromatography, coupled with one and two dimensional chromatography and protein sequencing of components of rabbit uterus and chick oviduct, including nontransformed progestin receptors. In addition, we will analyze cellular extracts of IM9, Jurkat, MCF-7 cells after hormonal and immunosuppressant treatment. Our second goal will be accomplished by binding assays (3H FK506, 3H ATP, etc.) of immunoprecipitated p59. Finally, after treating IM9, Jurkat, MCF, etc. cells with sex steroids, glucocorticoids, antihormones, and/or immunosuppressants we will study the cellular control of p59 by Western blotting, Northern blotting, and nuclear runoff transcription assays.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI034774-02
Application #
2069945
Study Section
Reproductive Endocrinology Study Section (REN)
Project Start
1993-03-01
Project End
1997-02-28
Budget Start
1994-03-01
Budget End
1995-02-28
Support Year
2
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Toledo
Department
Obstetrics & Gynecology
Type
Schools of Medicine
DUNS #
807418939
City
Toledo
State
OH
Country
United States
Zip Code
43614
Pang, H; Faber, L E (2001) Estrogen and rapamycin effects on cell cycle progression in T47D breast cancer cells. Breast Cancer Res Treat 70:21-6
Tai, P K; Albers, M W; McDonnell, D P et al. (1994) Potentiation of progesterone receptor-mediated transcription by the immunosuppressant FK506. Biochemistry 33:10666-71
Tai, P K; Chang, H; Albers, M W et al. (1993) P59 (FK506 binding protein 59) interaction with heat shock proteins is highly conserved and may involve proteins other than steroid receptors. Biochemistry 32:8842-7
Adams, P W; Pagel, E; Orosz, C G (1992) Detection of donor-reactive alloantibody in the early posttransplant period. Elimination of therapeutic ALG as a complicating factor. Transplantation 53:1302-5
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Orosz, C G; Zinn, N E; Sirinek, L P et al. (1988) In vivo mechanisms of alloreactivity--IV. Cyclosporine differentially impairs accumulation of donor-reactive CTL but not donor-reactive alloantibody in murine sponge matrix allografts. Int J Immunopharmacol 10:305-16
Orosz, C G; Adams, P W; Ferguson, R M (1987) Frequency of human alloantigen-reactive T lymphocytes. II. Method for limiting dilution analysis of alloantigen-reactive helper T cells in human peripheral blood. Transplantation 43:718-24
Orosz, C G; Zinn, N E; Sirinek, L et al. (1986) In vivo mechanisms of alloreactivity. I. Frequency of donor-reactive cytotoxic T lymphocytes in sponge matrix allografts. Transplantation 41:75-83
Orosz, C G; Zinn, N E; Sirinek, L P et al. (1986) In vivo mechanisms of alloreactivity. II. Allospecificity of cytotoxic T lymphocytes in sponge matrix allografts as determined by limiting dilution analysis. Transplantation 41:84-92

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