The Chemistry of the Hormones

Abstract

ADRENAL Venning et al. (1, 2) developed a method for the assay of urinary corticosteroid'like substances based upon the glycogen deposition in the livers of glucose treated adrenalectomized mice. A glycogenic unit is defined as the equivalent of the biological activity of 1 p.g. of 17-hydroxy-ll-dehydrocorticosterone. It was pointed out by these workers that the administration of glucose increased the sensitivity of the test many times. Similar assay methods have also been described by Dorfman et al. (3) and Eggleston et al. (4) using adrenalectomized mice without glucose treatment. Dorfman et al. (5) reexamined the cold test of Selye & Schenker (6) for the bioassay of urinary corticoids and found that the method was not practical for extended routine use.Two chemical methods for the determination of the urinary corticosteroid-like substances have also appeared. Heard & Sobel (7) introduced a color metric method, based on the reducing sugar like property of the adrenal cortical hormones, for the quantitative estimation of these steroids; the color was produced by the reduc tion of phosphomolybdic acid to molybdenum blue in glacial acetic acid. The reaction is given by steroids containing a primary or secondary (but not tertiary) , a-ketol grouping, an a, ,B-unsatu rated 3-ketone group, or both. Thus, desoxycorticosterone exhibits the strongest reducing properties. The method has applied to the estimation of urinary neutral lipide soluble reducing substances as an index of adrenal cortical function (8) and reveals a good agree ment between results obtained by this chemical method and that by the bioassay technique of Venning et al. (1). The method of Lowenstein et al. (9) in determining cortico steroids in urines is based on period ate oxidation of the primary alcohol group at C21 which yields one mole of formaldehyde per 1 This review includes the following subjects: adrenals, gonads, insulin, and anterior pituitary. Due to the limitation of space, other topics have been omitted. 291 Annu. Rev. Biochem. 1947.16:291-322. Downloaded from www.annualreviews.org Access provided by University of Sussex on 02/07/15. For personal use only. Quick links to online content Further ANNUAL REVIEWS

Studies on pituitary adrenocorticotrophin. 1. Ultrafiltration of the hormone

“…is itself ultrafilterable, makes it unlikely that there is more than one adrenocorticotrophic factor rmone by the (measured by adrenal hypertrophy and depletion of adrenal ascorbic acid) in the pituitary extracts, as suggested by Li (1947). Since a protein of molecular weight 20,000 cannot pass through the membranes used in this work, it seems probable that the protein <0-07 hormone can be dissociated under certain conditions <0-01…”

“…This conclusion is supported by the observations of Li et al (1943) who were able to hydrolyse 38 % of the protein hormone with pepsin without affecting the biological activity. Later Li (1947) partially hydrolysed the protein hormone with hydrochloric acid and found adrenocorticotrophic activity in the hydrolysates. Anselmino, Hoffmann & Herold (1934a, b) claimed to have ultrafiltered adrenocorticotrophin through a collodion membrane.…”

Physicochemical Characterization of Pituitary Follicle-Stimulating Hormone

“…3, the isoelectric point of the hormone is estimated to be at pH 4.5. It is of interest that this value is not greatly different from that of the interstitial cell-stimulating hormone (Li, 1947). Diffusion.--Experimental diffusion curves in one experiment were normalized and then compared with the ideal Gaussian curve.…”

“…It is not uncommon that the same hormone isolated from different species has different physicochemical characteristics. We have shown previously that the lactogenic hormones from ox and sheep glands (Li, 1947) may b~ differentiated by their tyrosine content and solubility characteristics. Similarly, interstitial cell-stimulating hormone preparations from hog and sheep glands appear to be different proteins with widely different molecular weights and isoelectric points (Li, 1947).…”

“…We have shown previously that the lactogenic hormones from ox and sheep glands (Li, 1947) may b~ differentiated by their tyrosine content and solubility characteristics. Similarly, interstitial cell-stimulating hormone preparations from hog and sheep glands appear to be different proteins with widely different molecular weights and isoelectric points (Li, 1947). In the case of insulin, Sanger (1949) has demonstrated a difference between ox and hog preparations in terminal end groups.…”