Author/Editor     Cassetta, Alberto; Buedefeld, Tomaž; Lanišnik-Rižner, Tea; Kristan, Katja; Stojan, Jure; Lamba, Doriano
Title     Crystallization, X-ray diffraction analysis and phasing of 17beta-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus
Type     članek
Source     Acta crystallographica section F
Vol. and No.     Letnik F61
Publication year     2005
Volume     str. 1032-4
ISSN     1744-3091
Language     eng
Abstract     17ß-Hydroxysteroid dehydrogenase from the filamentous fungus Cochlioholus lucuatus (17ß-HSDcI) is an NADP(H)-dependent enzyme that preferentially catalyses the oxidoreduction of oestrogens and androgens. The enzyme belongs to the short-chain dehydrogenase/reductase supertamily and is the only fungal hydroxysteroid dehydrogenase known to date. 77ß-HSDcI has recently been characterized and cloned and has been the subject of several functional studies. Although several hypotheses on the physiological role of 17ß-HSDcI in fungal metabolism have been formulated, its function is still unclear. An X-ray crystallographic study has been undertaken and the optimal conditions for crystallization of 17ß-HSDcI (apo form) were established, resulting in well shaped crystals that diffracted to 1.7A resolution. The space group was identified as 14 1 22, with unit-cell parameters a = b = 67.14, c= 266.77 A. Phasing was successfully performed by Patterson search techniques. A catalytic inactive mutant Tyr167Phe was also engineered, expressed, purified and crystallized for functional and structural studies.
Descriptors     ASCOMYCETES
17-HYDROXYSTEROID DEHYDROGENASES
RECOMBINANT PROTEINS
CRYSTALLIZATION
X-RAY DIFFRACTION