Dwain D. Hagerman

The hydrolysis of estrone sulfate by rat kidney microsomal sulfatase

Abstract A survey of the distribution of enzyme activity for the hydrolysis of estrone sulfate in various organs of the rat showed the kidney to be the most active. The hydrolytic activity is located primarily in the microsomal fraction of the kidney but could not be solubilized. The pH profile of the enzyme activity, its thermal stability, its substrate specificity, and its kinetic properties suggest that this enzyme is distinct from previously described kidney sulfatases.

Extraction of RNA from rat uterus

Abstract Methods for the extraction of RNA from the immature rat uterus have been critically evaluated. Primary emphasis was placed upon minimizing degradation of RNA. The procedure of choice consists of extraction with phenol and sodium dodecyl sulfate at 50° in the presence of bentonite, followed by deoxyribonuclease treatment of the extracted material. The product contains an average of 55% of the total RNA and >26% of the rapidly-labeled RNA (20-min pulse of [5- 3 H]uridine) of the uterus. (The recoveries are based on values obtained by hot HClO 4 extraction of RNA.) Almost all of the rapidly-labeled RNA in the product is of high molecular weight, with sedimentation coefficients in the range 30–50 S on sucrose gradients, which suggests that degradation of the material has been minimized. Estradiol given intraperitoneally to immature rats 2, 4, or 12 h before sacrifice did not affect the pattern of distribution of this high-molecular-weight, rapidly-labeled RNA (10-, 20-, or 40-min pulse of [5- 3 H]uridine) on sucrose gradients. Estradiol given intraperitoneally 20 min before sacrifice to rats ovariectomized at 28 days and used for experimentation 18–33 days after ovariectomy also did not affect the pattern of distribution of rapidly-labeled RNA (10-min pulse of [5- 3 H]uridine) on sucrose gradients.

Thin layer chromatography of some Δ5-3β-hydroxysteroids

Abstract The R F values for Δ 5 -pregnenolone, 17α-hydroxy-Δ 5 -pregnenolone, dehydroepiandrosterone, and 16-dehydro-Δ 5 -pregnenolone in 19 different solvent systems for thin layer chromatography on silica gel are given. A general method of determining the optimal solvent system for the separation of a group of compounds from a variety of possible systems is described.