title: Crystallization, X-ray diffraction analysis and phasing of 17beta-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus.
abstract: 17beta-Hydroxysteroid dehydrogenase from the filamentous fungus Cochliobolus lunatus (17beta-HSDcl) is an NADP(H)-dependent enzyme that preferentially catalyses the oxidoreduction of oestrogens and androgens. The enzyme belongs to the short-chain dehydrogenase/reductase superfamily and is the only fungal hydroxysteroid dehydrogenase known to date. 17beta-HSDcl has recently been characterized and cloned and has been the subject of several functional studies. Although several hypotheses on the physiological role of 17beta-HSDcl 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 17beta-HSDcl (apo form) were established, resulting in well shaped crystals that diffracted to 1.7 A resolution. The space group was identified as I4(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.
authors.affiliation: Institute of Crystallography, CNR, Trieste Outstation, Area Science Park-Basovizza, S.S.14, I-34012 Trieste, Italy. alberto.cassetta@ic.cnr.it
authors.forename: Alberto
authors.initials: A
authors.lastname: Cassetta
authors.forename: Tomaz
authors.initials: T
authors.lastname: Büdefeld
authors.forename: Tea Lanisnik
authors.initials: TL
authors.lastname: Rizner
authors.forename: Katja
authors.initials: K
authors.lastname: Kristan
authors.forename: Jure
authors.initials: J
authors.lastname: Stojan
authors.forename: Doriano
authors.initials: D
authors.lastname: Lamba
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000737:chemistry* / Q000235:genetics; D001203:Ascomycota / Q000201:enzymology*; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D014961:X-Ray Diffraction
chemical_list: D015067:17-Hydroxysteroid Dehydrogenases; C044256:3 (or 17)-beta-hydroxysteroid dehydrogenase