Crystallization and preliminary X-ray diffraction analysis of human adenovirus

Journal Article · Thu Jul 22 04:00:00 EDT 2010 · Virology

DOI:https://doi.org/10.1016/j.virol.2010.03.028· OSTI ID:1002533

Reddy, V S; Natchiar, S K; Gritton, L; Mullen, T -M; Stewart, P L; Nemerow, G R ^[1]

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1. Scripps

Replication-defective and conditionally replicating adenovirus (AdV) vectors are currently being utilized in {approx}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){sub 2}, 14% PEG 550 MME, 15% glycerol in 100 mM Tris-HCl 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{sup o}, {beta} = 91.71{sup o}, {gamma} = 118.08{sup o} 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.

• Research Organization: Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
• Sponsoring Organization: USDOE
• OSTI ID: 1002533
• Journal Information: Virology, Journal Name: Virology Journal Issue: (1) ; 06, 2010 Vol. 402; ISSN VIRLAX; ISSN 0042-6822
• Country of Publication: United States
• Language: ENGLISH

Related Subjects

43 PARTICLE ACCELERATORS
ADENOVIRUS
CLINICAL TRIALS
CRYSTALLIZATION
FIBERS
GENES
GLYCEROL
MODIFICATIONS
ORIENTATION
RESOLUTION
SPACE GROUPS
SYNCHROTRONS
VECTORS
X-RAY DIFFRACTION