TY - JOUR
T1 - Purification, crystallization and preliminary X-ray crystallographic analysis of 3-ketosteroid Δ 1-dehydrogenase from Rhodococcus erythropolis SQ1
AU - Rohman, Ali
AU - Van Oosterwijk, Niels
AU - Dijkstra, Bauke W.
PY - 2012/5
Y1 - 2012/5
N2 - 3-Ketosteroid Δ 1-dehydrogenase plays a crucial role in the early steps of steroid degradation by introducing a double bond between the C1 and C2 atoms of the A-ring of its 3-ketosteroid substrates. The 3-ketosteroid Δ 1-dehydrogenase from Rhodococcus erythropolis SQ1, a 56 kDa flavoprotein, was crystallized using the sitting-drop vapour-diffusion method at room temperature. The crystals grew in various buffers over a wide pH range (from pH 5.5 to 10.5), but the best crystallization condition consisted of 2%(v/v) PEG 400, 0.1 M HEPES pH 7.5, 2.0 M ammonium sulfate. A native crystal diffracted X-rays to 2.0 Å resolution. It belonged to the primitive orthorhombic space group P212121, with unit-cell parameters a = 107.4, b = 131.6, c = 363.2 Å, and contained eight molecules in the asymmetric unit. The initial structure of the enzyme was solved using multi-wavelength anomalous dispersion (MAD) data collected from a Pt-derivatized crystal.
AB - 3-Ketosteroid Δ 1-dehydrogenase plays a crucial role in the early steps of steroid degradation by introducing a double bond between the C1 and C2 atoms of the A-ring of its 3-ketosteroid substrates. The 3-ketosteroid Δ 1-dehydrogenase from Rhodococcus erythropolis SQ1, a 56 kDa flavoprotein, was crystallized using the sitting-drop vapour-diffusion method at room temperature. The crystals grew in various buffers over a wide pH range (from pH 5.5 to 10.5), but the best crystallization condition consisted of 2%(v/v) PEG 400, 0.1 M HEPES pH 7.5, 2.0 M ammonium sulfate. A native crystal diffracted X-rays to 2.0 Å resolution. It belonged to the primitive orthorhombic space group P212121, with unit-cell parameters a = 107.4, b = 131.6, c = 363.2 Å, and contained eight molecules in the asymmetric unit. The initial structure of the enzyme was solved using multi-wavelength anomalous dispersion (MAD) data collected from a Pt-derivatized crystal.
KW - 3-ketosteroid Δ -dehydrogenase
KW - Rhodococcus erythropolis SQ1
UR - http://www.scopus.com/inward/record.url?scp=84862576116&partnerID=8YFLogxK
U2 - 10.1107/S1744309112011025
DO - 10.1107/S1744309112011025
M3 - Article
C2 - 22691786
AN - SCOPUS:84862576116
SN - 1744-3091
VL - 68
SP - 551
EP - 556
JO - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
JF - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
IS - 5
ER -