STUDIES ON THE UPTAKE AND BINDING OF 5α-DIHYDROTESTOSTERONE BY RAT VENTRAL PROSTATE CELL NUCLEI IN VITRO

1970 ◽  
Vol 65 (3) ◽  
pp. 533-540 ◽  
Author(s):  
Olav Unhjem
Keyword(s):  
Sodium Chloride ◽  
Gel Filtration ◽  
Lower Degree ◽  
Ventral Prostate ◽  
Supernatant Fraction ◽  
Cell Nuclei ◽  
Gel Filtration Chromatography ◽  
Prostate Cell ◽  
Rat Ventral Prostate

ABSTRACT Following incubation of rat ventral prostate homogenates with 1,2-3H]-5α-dihydrotestosterone, the steroid was taken up and bound by the nuclei. Sodium chloride extraction of the nuclei removed macromolecules which might at least be partially responsible for the binding. Purified nuclei which were incubated with the radioactive steroid in either Tris-EDTA buffer or a 105 000 × g supernatant fraction prepared from the organ showed a lower degree of binding. Gel filtration chromatography of sodium chloride extracts from these nuclei disclosed binding of steroids to macromolecules. In all the experiments the steroids bound by nuclei consisted of unchanged 5α-dihydrotestosterone.

BINDING OF ANDROGEN TO COMPONENTS OF RAT VENTRAL PROSTATE NUCLEI IN VITRO

1970 ◽  
Vol 63 (1) ◽  
pp. 69-78 ◽  
Author(s):  
Olav Unhjem
Keyword(s):  
Sodium Chloride ◽  
Radioactive Material ◽  
Ventral Prostate ◽  
Major Portion ◽  
Nuclear Chromatin ◽  
Rat Ventral Prostate

ABSTRACT Following incubation of rat ventral prostate slices with [1,2-3H]testosterone in vitro, radioactive material is found to be associated with the nuclei. Sodium chloride extraction of the nuclei removes macromolecules which might at least partially be responsible for this association. These macromolecules are probably components of the nuclear chromatin. Analyses of the radioactive material indicate that the major portion consists of dihydrotestosterone* and to a lesser extent of testosterone. All the radioactive material is linked non-covalently to the nuclear components responsible for the association.

PARTIAL SEPARATION OF A 3α-KETOSTEROID OXIDOREDUCTASE AND AN ANDROGEN BINDING SUBSTANCE PRESENT IN RAT VENTRAL PROSTATE CYTOPLASM

1970 ◽  
Vol 65 (3) ◽  
pp. 525-532 ◽  
Author(s):  
Olav Unhjem
Keyword(s):  
Enzymatic Activity ◽  
Reduced Glutathione ◽  
Enzymatic Reaction ◽  
Gel Filtration ◽  
Ventral Prostate ◽  
Supernatant Fraction ◽  
Rat Ventral Prostate ◽  
Molecular Sizes ◽  
Androgen Binding ◽  
Fraction Addition

ABSTRACT The presence of a 3α-ketosteroid oxidoreductase in the rat ventral prostate cytoplasm has been demonstrated. The enzyme was confined to the 105 000 × g supernatant fraction with very little activity in the mitochondrial-microsomal fraction. Addition of NADPH2 was necessary for the enzymatic reaction to proceed. The enzymatic activity was lost when supernatant fractions were dialysed against 0.1 m Tris-HCl buffer, pH 7.4 or when chromatographed by gel filtration using the same buffer. When either L-cysteine, reduced glutathione, 2-mercaptoethanol or EDTA was added to the buffer, the enzymatic activity was preserved. The molecular weight of the enzyme was calculated to be 40 000–50 000. 3H-5α-dihydrotestosterone associates with macromolecules in the 105 000 × g supernatant fraction which are of two molecular sizes (60 000–70 000 and ∼ 200 000). These complexes could not be demonstrated by gel filtration using buffers containing either reduced glutathione, 2-mercaptoethanol or EDTA. By using L-cysteine, the small molecular complex was preserved.

LOCALIZATION OF AN ANDROGEN BINDING SUBSTANCE FROM THE RAT VENTRAL PROSTATE

1969 ◽  
Vol 60 (4) ◽  
pp. 571-578 ◽  
Author(s):  
Olav Unhjem ◽  
Kjell J. Tveter
Keyword(s):  
Gel Filtration ◽  
The Other ◽  
Male Rats ◽  
Ventral Prostate ◽  
Rat Ventral Prostate ◽  
Binding Substance ◽  
Liver And Kidney ◽  
Androgen Binding ◽  
Filtration Technique

ABSTRACT A gel filtration technique has been used for analysing the interaction of testosterone and/or its metabolites with macromolecules in the ventral prostate, rectus abdomonis muscle, liver and kidney from castrated adult male rats. Following administration of (1,2-3H)-testosterone, association of radioactivity with soluble macromolecules was demonstrated in the ventral prostate but not in the other organs analysed. A similar binding was observed when slices of ventral prostate were incubated in vitro with testosterone. When 105 000 × g supernatants from the ventral prostate were incubated with (1,2-3H)-testosterone at 37° C, radioactivity was also recovered associated with macromolecules. No association could be detected when the incubation was performed at 0° C.

FURTHER STUDIES ON A SOLUBLE ANDROGEN-MACROMOLECULAR COMPLEX FROM THE RAT VENTRAL PROSTATE

1970 ◽  
Vol 65 (3) ◽  
pp. 517-524 ◽  
Author(s):  
Olav Unhjem
Keyword(s):  
Hydrogen Atom ◽  
Keto Group ◽  
Ventral Prostate ◽  
Metabolic Inhibitors ◽  
Macromolecular Complex ◽  
Structural Requirements ◽  
Macromolecular Complexes ◽  
Rat Ventral Prostate

ABSTRACT The ability of various steroids and metabolic inhibitors to influence the binding of androgen to soluble macromolecules in the rat ventral prostate was evaluated in vitro. The results obtained revealed some structural requirements of steroids for binding to the macromolecules. An androstane skeleton with the α-configuration of the hydrogen atom at position 5 seemed to be essential for binding as well as a keto group at position 3. N-ethylmaleimide, Na-iodoacetate and p-hydroxymercuribenzoate inhibited the binding of androgen to macromolecules. The androgen-macromolecular complexes appeared to be rather stable at temperatures below 5°C.

scholarly journals Incorporation of Testosterone and 5α-Dihydrotestosterone into Rat Ventral Prostate in vitro

1970 ◽  
Vol 17 (6) ◽  
pp. 453-458 ◽  
Author(s):  
JUN SHIMAZAKI ◽  
JIN SATO ◽  
HISAKO NAGAI ◽  
KEIZO SHIDA
Keyword(s):  
Ventral Prostate ◽  
Rat Ventral Prostate

Cell surface modifications in the epithelium of rat ventral prostate during adaptation to in vitro conditions: An ultrastructural study

In Vitro ◽  
1984 ◽  
Vol 20 (3) ◽  
pp. 216-228 ◽  
Author(s):  
Frederick B. Merk ◽  
Paul W. L. Kwan ◽  
Stanley Spilman ◽  
Louis Terracio ◽  
William H. J. Douglas
Keyword(s):  
Cell Surface ◽  
Ultrastructural Study ◽  
Surface Modifications ◽  
Ventral Prostate ◽  
Rat Ventral Prostate

THE UPTAKE OF TESTOSTERONE AND ZINC IN VITRO BY THE HUMAN BENIGN HYPERTROPHIC PROSTATE

1973 ◽  
Vol 58 (3) ◽  
pp. 405-419 ◽  
Author(s):  
M. JOAN REED ◽  
S. R. STITCH
Keyword(s):  
Ion Exchange ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Zinc Binding ◽  
Supernatant Fraction ◽  
Low Molecular Weight ◽  
Molecular Weight Peak ◽  
Steroid Binding

SUMMARY The uptake of 65Zn and [1,2-3H]testosterone by minced tissue of human benign hypertrophic prostates and the subcellular distribution of radioactivity were examined. The nature of steroid and 65Zn binding by the cytosol (105000 g supernatant) fraction was investigated by gel filtration, ion-exchange chromatography and electrophoresis. It was found that steroid binding after incubation at 4°C was specific. One or two regions of steroid binding were observed after gel filtration of the cytosol using Sephadex G-200, depending upon incubation conditions. Binding of 65Zn was found in the low molecular weight peak after G-200 gel filtration. Equimolar CdCl2 and 65ZnCl2 were incubated with [1,2-3H]testosterone and minced tissue and the cytosol was subjected to gel filtration. Compared with control values, without CdCl2, reduction of 65Zn binding by about 50% occurred, while binding of 3H-labelled steroid was unaffected. Electrophoresis and ion-exchange chromatography showed that 65Zn and 3H-labelled steroid were bound to different proteins. A sample of the zinc-binding protein was prepared by ion-exchange chromatography and the homogeneity was checked by electrophoresis.

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