Crystallization and preliminary X-ray diffraction analysis of human adenovirus.
Replication-defective and conditionally replicating adenovirus (AdV) vectors are currently being utilized in approximately 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 100mM HEPES pH 7.0, 200mM Ca(OAc)(2), 14% PEG 550 MME, 15% glycerol in 100mM Tris-HCl 8.5. Freshly grown crystals diffracted well to 4.5A resolution and weakly to 3.5A at synchrotron sources. HAdV crystals belong to space group P1 with unit cell parameters a=854.03A, b=855.17A, c=865.24A, 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.
Reddy
V S
VS
The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. reddyv@scripps.edu
Natchiar
S K
SK
Gritton
L
L
Mullen
T-M
TM
Stewart
P L
PL
Nemerow
G R
GR
D000260:Adenoviruses, Human / Q000737:chemistry* / Q000648:ultrastructure*; D002021:Buffers; D018360:Crystallography, X-Ray; D006801:Humans
D002021:Buffers
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