The study of crystallization of estrogenic 17beta-hydroxysteroid dehydrogenase with DHEA and DHT at elevated temperature.
Most crystallization experiments of macromolecules are carried out at a constant temperature. Room temperature (22 degrees C) and 4 degrees C are the most widely used settings in crystallization. In practice, crystal growth at relatively high temperatures has often been avoided for macromolecular crystallization. Human estrogenic 17beta-hydroxysteroid dehydrogenase has been crystallized in complex with dehydroepiandrosterone or dihydrotestosterone. The crystallization experiments were carried out at 27 degrees C. The 17beta-HSD1 crystals were greatly improved at the elevated temperature. The effects of higher temperatures on crystal growth were studied. High temperatures stimulated the nucleation of 17beta-HSD1, increased the rate of crystal growth, and higher occupancy of substrates was obtained in the crystal structure. This method also reduced the formation of twin crystals. Since temperature is the easiest factor to control in the laboratory, crystallization at elevated temperatures provides an efficient method to improve protein crystal growth. The mechanism of the effect of temperature and relative techniques are discussed.
Han
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Medical Research Council Group in Molecular Endocrinology and Oncology, CHUL Research Center and Laval University, Ste-Foy, Quebec, G1V 4G2, Canada.
Lin
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D015067:17-Hydroxysteroid Dehydrogenases / Q000737:chemistry* / Q000378:metabolism*; D003460:Crystallization; D003687:Dehydroepiandrosterone / Q000737:chemistry / Q000378:metabolism*; D013196:Dihydrotestosterone / Q000737:chemistry / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D011485:Protein Binding; D015203:Reproducibility of Results; D014961:X-Ray Diffraction
D013196:Dihydrotestosterone; D003687:Dehydroepiandrosterone; D015067:17-Hydroxysteroid Dehydrogenases; C044256:3 (or 17)-beta-hydroxysteroid dehydrogenase