Methods for measuring ligand dissociation and nuclear receptor turnover in whole cells.
Understanding the molecular mechanisms of steroid hormone action requires assays that measure rates of ligand dissociation and receptor degradation. Ligand dissociation is a pseudo-first order reaction of a high affinity [3H]-labeled ligand. Receptor turnover as described here is the rate of degradation of a radiolabeled receptor. The methods make use of transient expression of a nuclear receptor in cultured cells and are applicable to all nuclear receptors. Rates of ligand dissociation and receptor degradation provided the first insight into the interdomain interactions of the androgen receptor and the molecular basis for the phenotypic effects of naturally occurring androgen receptor loss-of-function germline mutations and gain-of-function somatic mutations, and for the potency differences between the biologically active androgens, testosterone, and dihydrotestosterone.
Wilson
Elizabeth M
EM
Laboratories for Reproductive Biology, Department of Pediatrics, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, USA.
D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D006801:Humans; D008024:Ligands; D008297:Male; D011471:Prostatic Neoplasms / Q000378:metabolism; D011869:Radioligand Assay / Q000379:methods*; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D008024:Ligands; D011944:Receptors, Androgen; D018160:Receptors, Cytoplasmic and Nuclear