Involvement of a non-hormone-binding 90-kilodalton protein in the nontransformed 8S form of the rabbit uterus progesterone receptor
Nontransformed 8S progesterone receptor (8S-PR) was purified by hormone-specific affinity chromatography from rabbit uterine low-salt cytosol containing 20 mM molybdate. In the eluate obtained with radioactive progestin, sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) showed the presence of several bands, including three that corresponded to approx.90, approx.120, and approx.85-kDa proteins. None of these three proteins was found in the eluate of the affinity column when the molybdate-containing cytosol was chromatographed in the presence of nonradioactive progesterone (mock purification). Subsequent purification of the affinity eluate by DEAE-Sephacel chromatography gave a single radioactive receptor peak at 0.15 M KCl with a sedimentation coefficient of 8.5 S. Silver staining after SDS-PAGE revealed that this purified 8S-PR fraction contained mainly the 120-, 90-, and 85-kDa proteins. (/sup 3/H)R5020-labeled 8S-PR purified by DEAE-Sephacel column chromatography was UV irradiated, and after SDS-PAGE the 120- and 85-kDa proteins were revealed, but the 90-kDa protein was not. Further evidence for the presence of the 90-kDa non-hormone-binding protein in the purified molybdate-stabilized nontransformed 8S-PR structure was demonstrated. In the course of this work, it was verified that 0.3 M KCl added in cytosol in the absence of molybdate dissociated the 8S-PR complex, and purified 120- and 85-kDa progestin binding proteins were obtained by hormone-specific affinity chromatography of the salt-treated cytosol. In summer, the results demonstrated that, as for the nontransformed avian 8S-PR the nontransformed 8S form of the rabbit uterus PR includes a non-hormone-binding 90-kDa protein.
- Research Organization:
- Paris-South Univ. Medical School, Bicetre, France
- OSTI ID:
- 6527754
- Journal Information:
- Biochemistry; (United States), Vol. 25:21
- Country of Publication:
- United States
- Language:
- English
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PROGESTERONE
RADIORECEPTOR ASSAY
RECEPTORS
BIOCHEMISTRY
PURIFICATION
ELECTROPHORESIS
ION EXCHANGE CHROMATOGRAPHY
LIQUID COLUMN CHROMATOGRAPHY
MOLYBDATES
PROTEINS
RABBITS
TRITIUM COMPOUNDS
ULTRAVIOLET RADIATION
UTERUS
ANIMALS
BODY
CHEMISTRY
CHROMATOGRAPHY
ELECTROMAGNETIC RADIATION
FEMALE GENITALS
HORMONES
ISOTOPE APPLICATIONS
KETONES
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
MOLYBDENUM COMPOUNDS
ORGANIC COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PREGNANES
RADIATIONS
REFRACTORY METAL COMPOUNDS
SEPARATION PROCESSES
STEROID HORMONES
STEROIDS
TRACER TECHNIQUES
TRANSITION ELEMENT COMPOUNDS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques