Crystallization and preliminary X-ray diffraction studies of the human adenovirus serotype 2 proteinase with peptide cofactor.
Recombinant human adenovirus serotype 2 proteinase (both native and selenomethionine-substituted) has been crystallized in the presence of the serotype 12, 11-residue peptide cofactor. The crystals (space group P3(1)21 or P3(2)21, one molecule per asymmetric unit, a = b = 41.3 angstrum, c = 197.0 angstrum) grew in solutions containing 20-40% 2-methyl-2,4-pentanediol (MPD), 0.1-0.2 M sodium citrate, and 0.1 M sodium HEPES, pH 5.0-7.5. Diffraction data (84% complete to 2.2 angstrum resolution with Rmerge of 0.0335) have been measured from cryopreserved native enzyme crystals with the Argonne blue (1,024 x 1,024 pixel array) charge-coupled device detector at beamline X8C at the National Synchrotron Light Source (operated by Argonne National Laboratory's Structural Biology Center). Additionally, diffraction data from selenomethionine-substituted proteinase, 65% complete to 2.0 angstrum resolution with Rmerge values ranging 0.05-0.07, have been collected at three X-ray energies at and near the selenium absorption edge. We have determined three of the six selenium sites and are initiating a structure solution by the method of multiwavelength anomalous diffraction phasing.
Keefe
L J
LJ
Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory, Illinois 60439, USA.
Ginell
S L
SL
Westbrook
E M
EM
Anderson
C W
CW
D000260:Adenoviruses, Human / Q000145:classification / Q000201:enzymology*; D003001:Cloning, Molecular; D003067:Coenzymes / Q000737:chemistry; D018360:Crystallography, X-Ray; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D012703:Serotyping
D003067:Coenzymes; D010455:Peptides; D003546:Cysteine Endopeptidases; C091827:adenain