Crystallization and preliminary X-ray analysis of the complex of NADH and 3alpha-hydroxysteroid dehydrogenase from Pseudomonas sp. B-0831.
The NAD(P)(+)-dependent enzyme 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD) catalyzes the reversible interconversion of hydroxyl and oxo groups at position 3 of the steroid nucleus. The complex of NADH and 3alpha-HSD from Pseudomonas sp. B-0831 was crystallized by the hanging-drop vapour-diffusion method. Refinement of crystallization conditions with microseeding improved the quality of the X-ray diffraction data to a resolution of 1.8 A. The crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 62.46, b = 82.25, c = 86.57 A, and contained two molecules, reflecting dimer formation of 3alpha-HSD, in the asymmetric unit.
Kataoka
Sachiyo
S
Graduate School of Agriculture, Kyoto Prefectural University, 1-5 Hangi-cho, Shimogamo, Sakyo-ku, Kyoto, Kyoto 606-8522, Japan.
Nakamura
Shota
S
Ohkubo
Tadayasu
T
Ueda
Shigeru
S
Uchiyama
Susumu
S
Kobayashi
Yuji
Y
Oda
Masayuki
M
D015096:3-Hydroxysteroid Dehydrogenases / Q000737:chemistry*; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific) / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry; D003460:Crystallization; D019281:Dimerization; D009243:NAD / Q000737:chemistry*; D011549:Pseudomonas / Q000201:enzymology*; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D009243:NAD; D015096:3-Hydroxysteroid Dehydrogenases; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)