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), Journal Name: Biochemistry; (United States) Vol. 25:21; ISSN BICHA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BIOCHEMISTRY
BODY
CHEMISTRY
CHROMATOGRAPHY
ELECTROMAGNETIC RADIATION
ELECTROPHORESIS
FEMALE GENITALS
HORMONES
ION EXCHANGE CHROMATOGRAPHY
ISOTOPE APPLICATIONS
KETONES
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MEMBRANE PROTEINS
MOLYBDATES
MOLYBDENUM COMPOUNDS
ORGANIC COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PREGNANES
PROGESTERONE
PROTEINS
PURIFICATION
RABBITS
RADIATIONS
RADIORECEPTOR ASSAY
RECEPTORS
REFRACTORY METAL COMPOUNDS
SEPARATION PROCESSES
STEROID HORMONES
STEROIDS
TRACER TECHNIQUES
TRANSITION ELEMENT COMPOUNDS
TRITIUM COMPOUNDS
ULTRAVIOLET RADIATION
UTERUS
VERTEBRATES