Structure-function relationships of the vitamin D hormone receptor
Avian intestinal cytosoluble receptors for 1,25-dihydroxyvitamin D/sub 3/ (1,25(OH)/sub 2/D/sub 3/) were subjected to limited trypsin digestion, endogenous proteolytic action, as well as carboxypeptidase treatment, and the physical and functional properties of the resulting discrete polypeptide fragments were identified and contrasted with the native 1,25(OH)/sub 2/D/sub 3/ receptor. Resultant fragments were followed by tracing either radioactive 1,25(OH)/sub 2/D/sub 3/ or by probing with anti-receptor monoclonal antibodies. Two differentially trypsin-sensitive effects on the 1,25(OH)/sub 2/D/sub 3/ receptor were noted when fragments were detected by their ability to bind 1,25(OH)/sub 2/(/sup 3/H)D/sub 3/. Two hormone-bound fragments of 40 and 30 kDa were formed; neither bound to DNA-cellulose nor anti-receptor monoclonal antibodies. Immunoblot technology was used to show the disappearance of the 60 kDA receptor with increasing trypsin concentrations, paralleling the appearance of an immunoreactive 20 kDA fragment. The 20 kDA fragment did not bind hormone but was capable of interacting with DNA-cellulose in a fashion identical to that of the 60 kDA receptor. In contrast to the exogeneous effect of trypsin, incubation of chick intestinal cytosol resulted in the time-dependent formation of an endogenous protease-derived fragment of 45 kDa.
- Research Organization:
- Arizona Univ., Tucson (USA)
- OSTI ID:
- 7002221
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
RECEPTORS
STRUCTURE-ACTIVITY RELATIONSHIPS
VITAMIN D
BIOCHEMICAL REACTION KINETICS
BIRDS
INTESTINES
MONOCLONAL ANTIBODIES
TIME DEPENDENCE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TRYPSIN
ANIMALS
ANTIBODIES
BODY
DIGESTIVE SYSTEM
ENZYMES
GASTROINTESTINAL TRACT
HYDROLASES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MEMBRANE PROTEINS
ORGANIC COMPOUNDS
ORGANS
PEPTIDE HYDROLASES
PROTEINS
REACTION KINETICS
SERINE PROTEINASES
VERTEBRATES
VITAMINS
550201* - Biochemistry- Tracer Techniques