Crystallization and preliminary X-ray diffraction studies of a mammalian steroid dehydrogenase.
20 beta-Hydroxysteroid dehydrogenase from the cytosolic fraction of neonatal pig testis is a NADPH-dependent enzyme that catalyzes the reduction of the C-20 ketone of C21-steroids. It is 85% homologous in amino acid sequence to the human enzyme, carbonyl reductase. The enzyme has been crystallized from 36% saturated ammonium sulfate in 10 mM 2-[N-Morpholino]ethanesulfonic acid buffer. The size and the quality of nicely formed square bi-pyramidal crystals were improved by using a "seeding" technique. The crystals diffract X-rays to at least 2.5 A resolution. The space group is P4(1)2(1)2 (or P4(3)2(1)2) and the unit-cell dimensions are a = b = 58.53 A, c = 165.64 A. There is one molecule (M(r) = 30.5 kDa; 289 amino acid residues) in the asymmetric unit. An intensity data set to 2.5 A has been collected with an overall Rmerge of 6.6% for all reflections.
Ghosh
D
D
Medical Foundation of Buffalo, Inc., NY 14203.
Erman
M
M
Pangborn
W
W
Duax
W L
WL
Nakajin
S
S
Ohno
S
S
Shinoda
M
M
D000818:Animals; D003349:Cortisone Reductase / Q000737:chemistry*; D003460:Crystallization; D013552:Swine; D014961:X-Ray Diffraction
D003349:Cortisone Reductase