Crystallization and preliminary X-ray diffraction analysis of human adenovirus
Abstract
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.
- Authors:
-
- Scripps
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1002533
- Resource Type:
- Journal Article
- Journal Name:
- Virology
- Additional Journal Information:
- Journal Volume: 402; Journal Issue: (1) ; 06, 2010; Journal ID: ISSN 0042-6822
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 43 PARTICLE ACCELERATORS; ADENOVIRUS; CLINICAL TRIALS; CRYSTALLIZATION; FIBERS; GENES; GLYCEROL; MODIFICATIONS; ORIENTATION; RESOLUTION; SPACE GROUPS; SYNCHROTRONS; VECTORS; X-RAY DIFFRACTION
Citation Formats
Reddy, V S, Natchiar, S K, Gritton, L, Mullen, T -M, Stewart, P L, Nemerow, G R, and Vanderbilt). Crystallization and preliminary X-ray diffraction analysis of human adenovirus. United States: N. p., 2010.
Web. doi:10.1016/j.virol.2010.03.028.
Reddy, V S, Natchiar, S K, Gritton, L, Mullen, T -M, Stewart, P L, Nemerow, G R, & Vanderbilt). Crystallization and preliminary X-ray diffraction analysis of human adenovirus. United States. https://doi.org/10.1016/j.virol.2010.03.028
Reddy, V S, Natchiar, S K, Gritton, L, Mullen, T -M, Stewart, P L, Nemerow, G R, and Vanderbilt). 2010.
"Crystallization and preliminary X-ray diffraction analysis of human adenovirus". United States. https://doi.org/10.1016/j.virol.2010.03.028.
@article{osti_1002533,
title = {Crystallization and preliminary X-ray diffraction analysis of human adenovirus},
author = {Reddy, V S and Natchiar, S K and Gritton, L and Mullen, T -M and Stewart, P L and Nemerow, G R and Vanderbilt)},
abstractNote = {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.},
doi = {10.1016/j.virol.2010.03.028},
url = {https://www.osti.gov/biblio/1002533},
journal = {Virology},
issn = {0042-6822},
number = (1) ; 06, 2010,
volume = 402,
place = {United States},
year = {Thu Jul 22 00:00:00 EDT 2010},
month = {Thu Jul 22 00:00:00 EDT 2010}
}