Crystallization and preliminary X-ray diffraction analysis of human adenovirus

Replication-defective and conditionally replicating adenovirus (AdV) vectors are currently being utilized in similar to 25% of human gene transfer clinical trials. Unfortunately, progress in vector development has been hindered by a lack of accurate structural information. Here we describe the crystallization and preliminary X-ray diffraction analysis of a HAdV5 vector that displays a short flexible fiber derived from HAdV35. Crystals of Ad35F were grown in 100 mM HEPES pH 7.0, 200 mM Ca(OAc)(2), 14% PEG 550 MME, 15% glycerol in 100 mM Tris-HCI 8.5. Freshly grown crystals diffracted well to 4.5 angstrom resolution and weakly to 3.5 angstrom at synchrotron sources. HAdV crystals belong to space group P1 with unit cell parameters a = 854.03 angstrom, b = 855.17 angstrom, c = 865.24 angstrom, alpha= 119.57 degrees. beta= 91.71 degrees, gamma= 118.08 degrees with a single particle in the unit cell. Self-rotation and locked-rotation function analysis allowed the determination of the particle orientation. Molecular replacement, density modification and phase-extension procedures are being employed for structure determination. (C) 2010 Elsevier Inc. All rights reserved.