Studies of molecular species of the human androgen receptor (AR): comparison of the physicochemical properties of the (/sup 3/H)methyltrienolone-AR complex formed in cytosol to the complex produced in intact genital skin fibroblasts

Journal Article · Tue Jul 01 00:00:00 EDT 1986 · J. Clin. Endocrinol. Metab.; (United States)

DOI:https://doi.org/10.1210/jcem-63-1-222· OSTI ID:5327238

Keenan, B S; Greger, N C; Hedge, A M

Two forms of the human genital skin fibroblast (GSF) androgen receptor (AR) complexed with (/sup 3/H)17 alpha-methyltrienolone were compared: 1) the intact complex formed in cytosol at 4 C (broken cell or B/C complex); and 2) the complex formed in the whole cell at 37 C (W/C complex). The intact form of the B/C complex was distinguished from partly degraded forms by the gel filtration profile in 0.5 M KCl. The W/C complex was considered to represent the transformed state of the receptor. The W/C complex had a smaller molecular radius than the B/C complex by gel filtration (Kav = 0.26-0.28 vs. 0.11-0.18). By low salt density gradient centrifugation, the B/C complex sedimented at 8.8S and the W/C complex at 6.6S. However, in 0.5 M KCl, each sedimented at 5.1S, and they were homogeneous, indicating that the monomeric forms differed markedly in molecular radius, but by only about 20,000 daltons in calculated mol wt (134,500 vs. 114,300 daltons). The complexes were separated from DNA, desalted, and compared by chromatography on DEAE-Sephacel and hydroxylapatite (HAP). The B/C complex bound readily to both column matrices and eluted from each as a sharp homogeneous peak: from DEAE at 172-190 mM KCl and from HAP at 123 mM phosphate. The W/C complex, however, was heterogeneous. One component did not bind to DEAE, and one eluted at 22-40 mM KCl. The W/C complex eluted from HAP as a peak at 42 mM, with a shoulder at 102 mM phosphate. Thus, transformation of the human genital skin fibroblast androgen receptor involves a major decrease in molecular radius and loss of negative charge with a possible loss of a 20,000-dalton macromolecular component.

• Research Organization: Baylor College of Medicine, Houston, TX
• OSTI ID: 5327238
• Journal Information: J. Clin. Endocrinol. Metab.; (United States), Vol. 1
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ANDROGENS
RECEPTORS
CHEMICAL PROPERTIES
FIBROBLASTS
MAN
MOLECULAR WEIGHT
SKIN
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANDROSTANES
ANIMAL CELLS
ANIMALS
BODY
CONNECTIVE TISSUE CELLS
HORMONES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
ORGANIC COMPOUNDS
ORGANS
PRIMATES
PROTEINS
SOMATIC CELLS
STEROID HORMONES
STEROIDS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques