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, presen...

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Bibliographic Details
Published in:Protein science 1996-09, Vol.5 (9), p.1931-1933
Main Authors: Guarnéa, Alba, Verdaguer, Núaria, Fita, Ignasi, Kirchweger, Regina, Liebig, Hans‐Dieter, Blaas, Dieter, Skern, Tim
Format: Article
Language:English
Subjects:
Aphthovirus - enzymology
Crystallization
cysteine‐proteinase
Endopeptidases - analysis
Endopeptidases - chemistry
Endopeptidases - genetics
foot‐and‐mouth disease virus
Magnesium Chloride
Mutation
Osmolar Concentration
papain
Polyethylene Glycols
Recombinant Proteins - chemistry
Temperature
viral proteinase
X-Ray Diffraction
X‐ray crystallography
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Summary: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 Å and diffracted to, at most, 5 Å 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 × 0.3 × 0.7 mm3, diffracted to approximately 4.0 Å resolution, but presented a strong anisotropic mosaicity around the longest crystal axis. Crystals with a needlelike morphology and reaching sizes of about 0.2 × 0.3 × 1.2 mm3 diffracted beyond 3.5 Å 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 I212121) and unit cell dimensions a = 65.9 Å, b = 104.3 Å, and c = 124.0 Å, 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%.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.5560050921