Crystallization and preliminary X-ray diffraction studies of the Lb proteinase from foot-and-mouth disease virus.
Different crystal forms of the C23A mutant from the leader proteinase of foot-and-mouth disease virus were obtained by the hanging drop vapor diffusion technique, using MgCl2 and PEG 6000 as precipitants. Well-developed crystals, with cubic morphology growing to approximately 1.0 mm3 in size, presented a large unit cell parameter of 274.5 A and diffracted to, at most, 5 A resolution. A second type of crystal had a tetragonal appearance and these were obtained in droplets soaked in a silica gel matrix. These crystals, with an approximate size of 0.3 X 0.3 X 0.7 mm3, diffracted to approximately 4.0 A resolution, but presented a strong anisotropic mosaicity around the longest crystal axis. Crystals with a needlelike morphology and reaching sizes of about 0.2 X 0.3 X 1.2 mm3 diffracted beyond 3.5 A resolution and were stable to X-ray radiation for approximately one day when using a conventional source at room temperature. These crystals are orthorhombic with space group I222 (or I2(1)2(1)2(1)) and unit cell dimensions a = 65.9 A, b = 104.3 A, and c = 124.0 A, and appear well suited for high-resolution studies. Density packing considerations are consistent with the presence of two molecules in the asymmetric unit and a solvent content of approximately 54%.
Guarné
A
A
Centre d'Investigació i Desenvolupament (CSIC), Barcelona, Spain.
Kirchweger
R
R
Verdaguer
N
N
Liebig
H D
HD
Blaas
D
D
Skern
T
T
Fita
I
I
D005537:Aphthovirus / Q000201:enzymology*; D003460:Crystallization; D010450:Endopeptidases / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D015636:Magnesium Chloride; D009154:Mutation; D009994:Osmolar Concentration; D011092:Polyethylene Glycols; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; D015636:Magnesium Chloride; D011092:Polyethylene Glycols; D010450:Endopeptidases; C088747:leader proteinase, foot-and-mouth disease virus