Crystallization of a ternary complex of lactate dehydrogenase from Bacillus stearothermophilus.
Bacillus stearothermophilus lactate dehydrogenase was purified from an overexpressing Escherichia coli cell line. The enzyme has been crystallized in several different forms. All of these crystal forms were grown in the presence of NADH, sodium oxamate and fructose 1,6-bisphosphate. Three crystal forms have been characterized, an orthorhombic P2(1)2(1)2 (type III, a = 86 A, b = 105 A, c = 136 A) and two monoclinic P21 forms (type IV, a = 85 A, b = 118 A, c = 136 A, beta = 96 degrees; type V, a = 112 A, b = 85 A, c = 136 A, beta = 91 degrees). Precession photographs from these crystal forms are very alike, suggesting the molecular packing to be similar in all three forms. The P21 type IV crystals diffract to beyond 2 A spacing and are stable to irradiation with X-rays. A complete medium-resolution (4.7 A) dataset has been collected from a single crystal using synchrotron radiation. Rotation function studies with these data show the two tetramers of the asymmetric unit to be in very similar orientations. Higher-resolution data are being collected.
Wigley
D B
DB
Department of Biochemistry University of Bristol Medical School, England.
Muirhead
H
H
Gamblin
S J
SJ
Holbrook
J J
JJ
D002460:Cell Line; D004926:Escherichia coli; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D007770:L-Lactate Dehydrogenase; D046911:Macromolecular Substances; D014961:X-Ray Diffraction
D046911:Macromolecular Substances; D007770:L-Lactate Dehydrogenase