By Ulrich Westphal (auth.)
This is the tale of steroid-protein interactions as one investigator sees it. Following the final thought of this monograph sequence, it emphasizes the consequences and interpretations received within the author's laboratory, and is for this reason a subjective account. spotting, how ever, that the dialogue of the topic will be incomplete if the cloth have been constrained to at least one scientist's paintings, the basic in attaining ments of alternative investigators were integrated. An attempt has therefore been made to provide a balanced presentation and to permit the reader to determine in point of view the numerous aspects of the interactions among steroids and proteins. due to the fact this can be the 1st entire remedy of the subject, it kind of feels applicable to visit the roots, and check out to determine the way it started. the 1st chapters, for that reason, take the reader to the laboratories of these who very early conceived the importance of the attachment of dyes, medicines, and different conspicuous molecules to these colloids referred to as proteins. the invention of the steroid hormones set the degree for significant research in their interplay with proteins of assorted origins and capabilities - a technique that's carrying on with at the present time with expanding vigor.
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This can be the tale of steroid-protein interactions as one investigator sees it. Following the final thought of this monograph sequence, it emphasizes the consequences and interpretations received within the author's laboratory, and is accordingly a subjective account. spotting, how ever, that the dialogue of the topic will be incomplete if the cloth have been restricted to 1 scientist's paintings, the fundamental in attaining ments of alternative investigators were included.
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Extra info for Steroid-Protein Interactions
DE, measure the lowering of E as percentage of the values observed in the absence of protein. Fig. IV-5 indicates that they may be obtained directly from the absorbance values, without transforming them to molar extinction coefficients. d 4-3-ketosteroids to a protein (WESTPHAL and ASHLEY, 1958 a, 1959, 1962). dE is determined as a relatively small difference in absorbance in the pres- 38 IV. ground absorption by the protein, it is necessary that all experimental conditions are strictly identical, if the comparison of LIe obtained from different steroids is to be valid.
Differences in adsorption affinity may be utilized for separation of chemically related steroids; this has been shown by VAN BAELEN et al. (1967 a, b) who separated estradiol, estrone and estriol by gel filtration using Sephadex G-25 and Sephadex LH-20. Similar divergence of K' from theory has been found with other compounds of various chemical classes when studied in their adsorption behavior toward cross-linked dextrans (GELOTTE, 1960; MARSDEN, 1965; JANSON, 1967). PEARLMAN and CREPY (1967) have used the gel equilibration procedure to measure testosterone binding to BSA and HSA, and obtained association constants similar to those found in other laboratories.
S--- - - - - - - ---+ 15'(27000 x g) +- - - - - - - - - - - - - - - - S. XO' P I I . ------'---. _ _ _ _ P I SUPERNATANT I MICROSOMES Fig. IV-4. Equilibrium fractionation of rat liver cells. P, precipitate; S, supernatant. From DE VENUTO et al. (1962) 3' 36 IV. Basic Methods for Determination of Steroid-Protein Interactions contamination; this requires washing procedures which are ordinarily carried out by introducing fresh solutions at given stages into the fractionation system. Since the non-covalent interactions are regulated by equilibria of binding, a certain unbound percentage of the total steroid is removed by dissociation from its binding site with each addition of fresh wash medium.