Crystallization and preliminary X-ray diffraction studies of the Lb proteinase from foot-and-mouth disease virus

被引:7
作者
Guarne, A
Kirchweger, R
Verdaguer, N
Liebig, HD
Blaas, D
Skern, T
Fita, I
机构
[1] CSIC,CTR INVEST & DESENVOLUPAMENT,BARCELONA 08034,SPAIN
[2] UNIV VIENNA,FAC MED,INST BIOCHEM,A-1030 VIENNA,AUSTRIA
关键词
cysteine-proteinase; foot-and-mouth disease virus; papain; viral proteinase; X-ray crystallography;
D O I
10.1002/pro.5560050921
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 mm(3) in size, presented a large unit cell parameter of 274.5 Angstrom and diffracted to, at most, 5 Angstrom 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 mm(3), diffracted to approximately 4.0 Angstrom 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 mm(3) diffracted beyond 3.5 Angstrom 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 1222 (or I2(1)2(1)2(1)) and unit cell dimensions a = 65.9 Angstrom, b = 104.3 Angstrom, and c = 124.0 Angstrom, 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%.
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页码:1931 / 1933
页数:3
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