Sexual hormones in Achyla. V. Properties of hormone A of Achyla bisexualis

1. The hormonal coordinating mechanism of the sexual process in Achlya is briefly reviewed. 2. A technique is described for culturing the female plant of Achlya bisexualis in sufficient quantity to furnish material for the chemical study of hormone A. 3. A modification of the biological assay for hormone A is described. 4. Many of the properties of hormone A have been determined: (a) solubilities in common organic solvents, (b) adsorption, (c) stability, (d) inactivation, and (e) reactions with certain reagents. 5. A procedure is described whereby enormous enrichment of the active principle has been achieved

Xanthurenic acid and its rôle in the trytophane metabolism of pyroxidine-deficient rats

In a previous publication (1) the isolation of a green pigment from the urine of pyridoxine-deficient rats was described. The green pigment was shown to be the product of a reaction between ferric ammonium sulfate or other ferric salts and a compound whose nature was unknown. This compound has now been isolated in crystalline form. It is a yellow pigment and has been identified as xanthurenic acid

A new method for the separation of androgens from estrogens and for the partition of estriol from the estrone-estradiol fraction: with special reference to the identification and quantitative microdetermination of estrogens by ultraviolet absorption spectrophotometry

It is recognized generally that a qualitative and quantitative knowledge of the excretion pattern of the urinary estrogens is one index to an understanding of the functional activity of the ovary and adrenal cortex. Obviously, such determinations may be useful also in evaluating the normal and abnormal functions of other physiologically related endocrine glands as well as of organs like the liver and kidneys. The clinical applications of these data are self-evident. Various attempts have been made to circumvent the notoriously inaccurate values which have been obtained for the urinary estrogens by a variety of bioassay methods and calorimetric techniques (1, 2). The acknowledged shortcomings of these methods have led us to investigate the application of ultraviolet absorption spectrophotometry to the quantitative determination of the urinary estrogens in an attempt to develop an objective physical method for their accurate determination. It is known that the infra-red portion of the spectrum yields more differentially characteristic curves, but those of the ultraviolet range are more readily obtainable, and consequently better adapted to clinical use. This communication is concerned with studies of the following aspects of the problem: (1) spectrophotometric identification and quantitative micro determination of crystalline estrogens; (2) detection by spectrophotometric assay of gross errors in current methods for extraction and partition of estrogens; (3) studies on the ultraviolet absorption of substances comprising the background material; (4) separation of the phenolic estrogens from the so called neutral steroid fraction; (5) separation of urinary estrogens from other urinary phenolic substances by steam distillation; (6) micro-Girard separation of estrone from estradiol; (7) an essentially new method for the extraction and partition of crystalline estrone, estradiol, and estriol, and their quantitative assay by ultraviolet spectrophotometry

The incorporation of labeled lysine into the proteins of guinea pig liver homogenate

When C14-labeled lysine is incubated with guinea pig liver homogenate, α-aminoadipic, α-ketoadipic, and glutaric acids are formed from the lysine (1). These transformations were established by finding the radioactivity of the C14 tracer in the metabolic products. The homogenate proteins coagulated by boiling at pH 5 also contained radioactivity. The counts given by the proteins corresponded to about 0.02 to 0.03 per cent of that added as lysine; the extent of lysine incorporation into the proteins was of the same order of magnitude as Melchior and Tarver (2) had found after incubating S35-labeled methionine and Winnick et al. (3, 4) C14-labeled glycine with rat tissue homogenates. Yet we could not satisfy ourselves that the radioactivity remaining in the proteins in our experiments, although it persisted through exhaustive extraction, did not come from traces of adsorbed radioactive lysine. Some counts were found in the protein when the homogenate was boiled prior to incubation with isotopic lysine

Isolation of a peptide in guinea pig liver homogenate and its turnover of leucine

Leucine was synthesized with C14 in the carboxyl group. 10 mg. of the radioactive amino acid (DL) and 0.66 gm. (wet weight) of guinea pig liver homogenate were added to a reaction mixture containing 1.3 per cent of an amino acid mixture corresponding to the composition of casein and 0.005 M fumarate, all in a final volume of 4 ml. of isotonic saline solution(1) at pH 7.4. The reaction was carried out under oxygen for 6 hours at 38°

Incorporation in vitro of labeled amino acids into proteins of rabbit reticuloytes

Continuing our work on the incorporation of labeled amino acids into proteins (1), we have begun a study of the incorporation in vitro of C14-labeled glycine, L-histidine, L-leucine, and L-lysine into the proteins of rabbit reticulocytes. In preliminary experiments the incorporation into the hemoglobin isolated from the reticulocytes was determined. But, after it was found that plasma contains factors accelerating amino acid incorporation, it was decided to proceed as rapidly as possible toward the identification of these factors; we have, therefore, measured incorporation into the total proteins of the reticulocytes, since isolation of the hemoglobin was time-consuming. The results obtained with hemoglobin and with the total proteins are essentially the same, indicating that the other proteins of the reticulocytes incorporate amino acids at approximately the same rate as hemoglobin

Alpha-aminoadipic acid: A product of lysine metabolism

As part of a study of protein and peptide metabolism lysine was synthesized with C14 in the ε position and resolved into the L and D isomers. 10 mg. of labeled lysine dihydrochloride (either L- or D-) and 0.66 gm. (wet weight) of guinea pig liver homogenate were added to a reaction mixture containing 1.3 per cent of an amino acid mixture corresponding to the composition of casein except for lysine and 0.01 M α-ketoglutarate, all in a final volume of 4 ml. of isotonic saline solution.(1) The reaction was carried out under oxygen for 6 hours at 38°

Incorporation in vitro of labeled amino acids into bone marrow cell proteins

Nearly all experiments on the incorporation of labeled amino acids into tissue proteins in vitro have been done on tissues whose cell structure has been partially or completely disintegrated, e.g. tissue slices, segments, or homogenates. Since cell destruction reduces or abolishes the uptake of labeled amino acids (1), it seemed worth while to carry out studies on intact cells in vitro. Bone marrow cells were found to be suitable for this purpose. The labeled amino acids used were glycine-1-C14, L-leucine-1-C14, L-lysine-1-C14, and L-lysine-6-C14

The degradation of L-lysine in guinea pig liver homogenate: formation of alpha-aminoadipic acid

A summary of the little that is known of the metabolism of lysine in animals is as follows: it is indispensable in the diet, its α-amino group does not participate in reversible transamination reaction in vivo (2), neither the L nor D form is attacked by the appropriate amino acid oxidase, certain ε-nitrogen-substituted derivatives can replace lysine in the diet and their α-amino groups are oxidized by amino acid oxidases (3, 4), no α-nitrogen-substituted derivatives yet prepared can substitute for lysine in the diet (4-6)

The uptake in vitro of C14-labeled glycine, L-leucine, and L-lysine by different components of guinea pig liver homogenate

We have reported (1) that L-lysine labeled with C14 can be incorporated into the proteins of guinea pig liver homogenate under two different conditions. In the one case the enzyme used was the whole homogenate, the optimum pH was near 6.2, there was an obligatory requirement of calcium, and the incorporation was independent of oxygen. This set of conditions is designated below as the “acid calcium” condition. In the other case the enzyme system was the precipitate obtained by centrifuging the homogenate diluted 15-fold with Ringer’s solution at 2500 X g, the optimum pH was near to 7.3, the reaction was accelerated a little by calcium but the presence of calcium was not obligatory, and the incorporation was a little less under nitrogen than under oxygen. This set of conditions is designated below as the “alkaline” condition