Crystallization and preliminary x-ray diffraction studies of hyperthermostable glutamate dehydrogenase from Thermococcus profundus.
Recombinant glutamate dehydrogenase from a hyperthermophilic archaeon, Thermococcus profundus, was crystallized in the presence of both polyethylene glycol 8000 and lithium sulfate. Four types of crystals having different morphologies appeared in the crystallization trials; however, only one type was suitable for X-ray crystal structure analysis. The crystal belonged to the monoclinic space group P2(1) and the unit-cell parameters were a = 112.99, b = 163.70, c = 133.07 A, beta = 113.46 degrees at 110 K. The calculated V(M) value of 3.42 A(3) Da(-1) was acceptable when one hexamer of the enzyme, which was the physiological functional unit, occupied a crystallographic asymmetric unit. X-ray diffraction intensity data were collected to a resolution of 2.25 A with good statistics at the BL44B2 beamline of SPring-8.
Higuchi
S
S
The Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako, Saitama 351-0198, Japan.
Nakasako
M
M
Kudo
T
T
D003460:Crystallization; D018360:Crystallography, X-Ray; D004795:Enzyme Stability; D005969:Glutamate Dehydrogenase / Q000737:chemistry*; D011092:Polyethylene Glycols; D011994:Recombinant Proteins / Q000737:chemistry; D019712:Thermococcus
D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D005969:Glutamate Dehydrogenase