Steroid-Protein Interactions by Ulrich Westphal (auth.)

By Ulrich Westphal (auth.)

This is the tale of steroid-protein interactions as one investigator sees it. Following the overall idea of this monograph sequence, it emphasizes the consequences and interpretations received within the author's laboratory, and is hence a subjective account. spotting, how­ ever, that the dialogue of the topic will be incomplete if the fabric have been constrained to 1 scientist's paintings, the fundamental in attaining­ ments of different investigators were included. An attempt has hence been made to offer a balanced presentation and to let the reader to work out in standpoint the various features of the interactions among steroids and proteins. on the grounds that this is often the 1st finished therapy of the subject, it sort of feels acceptable to visit the roots, and take a look at to determine the way it begun. the 1st chapters, for this reason, take the reader to the laboratories of these who very early conceived the importance of the attachment of dyes, medicinal drugs, and different conspicuous molecules to these colloids referred to as proteins. the invention of the steroid hormones set the level for significant research in their interplay with proteins of assorted origins and features - a procedure that is carrying on with this present day with expanding vigor.

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This limitation has to be considered in the application of the original gel equilibration technique. GUERIGUIAN and CREPY (1969) have used the same principle to develop a new procedure to measure the concentration of the testosterone-binding fi-globulin in serum. In spite of the limitations inherent in this novel procedure, it may prove useful for the rapid assessment of protein binding to steroids, especially for comparative estimations under closely similar conditions. PEARLMAN (1970) has recently summarized the essential features of the technique, including calculations.

48 IV. Basic Methods for Determination of Steroid-Protein Interactions Whereas most steroids under usual experimental conditions are relatively stable substances, organic compounds labeled with radioisotopes have a general tendency to decompose on storage (BAYLY and EVANS, 1966, 1967). This is the result of self-irradiation which produces molecular alterations and, especially in the presence of oxygen, chemical reactions similar to those obtained with external ionizing radiation. , 1953). The extent of decomposition ranged from 1 to over 600/0 for various compounds; cholesterol over a period of about 18 months showed approximately 40010 destruction, based on the amount no longer precipitable with digitonin.

8. Turbidimetry An elegant refinement of the solubility method has been described by THERRIAULT and TAYLOR (1964) who determined the insoluble portion of the steroid by turbidimetric analysis; the procedure was developed in a study of phosphatidylserine interaction with HSA. In the earlier work of BISCHOFF, the insoluble steroid phase was separated from the equilibrated steroid-protein solution by filtration, and the protein-bound steroid was determined in the clear filtrate by 30 IV. Basic Methods for Determination of Steroid-Protein Interactions colorimetric procedures, following precipitation of the protein by ethanol.

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