Crystallization and preliminary X-ray diffraction analysis of naphthalene dioxygenase from Rhodococcus sp. strain NCIMB 12038.
The three-component naphthalene dioxygenase (NDO) enzyme system carries out the first step in the aerobic degradation of naphthalene to (+)-cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene by Rhodococcus sp. strain NCIMB 12038. The terminal oxygenase component (naphthalene 1,2-dioxygenase) that catalyzes this reaction belongs to the aromatic ring hydroxylating dioxygenase family and has been crystallized. These enzymes utilize a mononuclear non-heme iron centre to catalyze the addition of dioxygen to their respective substrates. In this reaction, two electrons, two protons and a dioxygen molecule are consumed. The Rhodococcus enzyme has only 33 and 29% sequence identity to the corresponding alpha- and beta-subunits of the NDO system of Pseudomonas putida NCIMB 9816-4, for which the tertiary structure has been reported. In order to determine the three-dimensional structure of the Rhodococcus NDO, diffraction-quality crystals have been prepared by the hanging-drop method. The crystals belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 87.5, b = 144, c = 185.6 A, alpha = beta = gamma = 90 degrees, and diffract to 2.3 A resolution.
Malik
Zulfiqar A
ZA
Department of Biochemistry, The University of Iowa, Iowa City, IA 52242, USA.
Allen
Christopher C R
CC
Gakhar
Lokesh
L
Lipscomb
David A
DA
Larkin
Michael J
MJ
Ramaswamy
S
S
D003460:Crystallization; D018360:Crystallography, X-Ray; D049308:Dioxygenases; D009097:Multienzyme Complexes / Q000737:chemistry*; D010105:Oxygenases / Q000737:chemistry*; D011487:Protein Conformation; D012240:Rhodococcus / Q000201:enzymology*
D009097:Multienzyme Complexes; D010105:Oxygenases; D049308:Dioxygenases; C038703:naphthalene dioxygenase