Crystallization and preliminary X‐ray diffraction studies of the human adenovirus serotype 2 proteinase with peptide cofactor

• Published in: Protein science Vol. 4; no. 8; pp. 1658 - 1660
• Main Authors: Keefe, Lisa J., Ginell, Stephan L., Westbrook, Edwin M., Anderson, Carl W.
• Format: Journal Article
• Language: English
• Published: Bristol Cold Spring Harbor Laboratory Press 01-08-1995
• Subjects: Adenoviruses, Human - classification; Adenoviruses, Human - enzymology; Cloning, Molecular; Coenzymes - chemistry; cryocrystallography; crystallization; Crystallography, X-Ray; Cysteine Endopeptidases - chemistry; Cysteine Endopeptidases - genetics; cysteine proteinase; Escherichia coli - genetics; Peptides - chemistry; Protein Conformation; proteinase/cofactor complex; selenomethionine; Serotyping; thiol proteinase; X‐ray crystallography
• ISSN: 0961-8368; 1469-896X
• DOI: 10.1002/pro.5560040826

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 P3121 or P3221, one molecule per asymmetric unit, a = b = 41.3 Å, c = 197.0 Å) 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 Å resolution with Rmerge of 0.0335) have been measured from cryopreserved native enzyme crystals with the Argonne blue (1,024 × 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 Å 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.