Crystallization and preliminary X-ray diffraction analysis of human adenovirus

Replication-defective and conditionally replicating adenovirus (AdV) vectors are currently being utilized in ∼ 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–HCl 8.5. Freshly grown crystals diffracted well to 4.5 Å resolution and weakly to 3.5 Å at synchrotron sources. HAdV crystals belong to space group P1 with unit cell parameters a = 854.03 Å, b = 855.17 Å, c = 865.24 Å, α = 119.57°, β = 91.71°, γ = 118.08° 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.