Crystallization, X-ray diffraction analysis and phasing of 17β-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 61; no. 12; pp. 1032 - 1034
• Main Authors: Cassetta, Alberto, Büdefeld, Tomaž, Lanišnik Rižner, Tea, Kristan, Katja, Stojan, Jure, Lamba, Doriano
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.12.2005; International Union of Crystallography
• Subjects: 17β-hydroxysteroid dehydrogenase; Cochliobolus lunatus; CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; Crystallization Communications; CRYSTALS; HSD; RESOLUTION; SDR; SPACE GROUPS; X-RAY DIFFRACTION
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309105034949

17β‐Hydroxysteroid dehydrogenase from the filamentous fungus Cochliobolus lunatus (17β‐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. 17β‐HSDcl has recently been characterized and cloned and has been the subject of several functional studies. Although several hypotheses on the physiological role of 17β‐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 17β‐HSDcl (apo form) were established, resulting in well shaped crystals that diffracted to 1.7 Å resolution. The space group was identified as I4122, with unit‐cell parameters a = b = 67.14, c = 266.77 Å. 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.