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4) Symposium Posters | Main^

P9. E0049: Structure Determination of Adeno-associated Virus 2 in P1 Cell with Three Complete Virus Particles.

2003 American Crystallographi Association Meeting, July 26-31, Covington, KY.
Qing Xie*,†, Thayumanasamy Somasundaram†, Smita Bhatia, Weishu Bu†, and Michael S. Chapman
*Dept. of Chemistry & Biochemistry, †Inst. of Molecular Biophysics, Kasha Laboratory, Florida State Univ., Tallahassee, FL 32306-4380, USA.

The atomic structure of adeno-associated virus 2 (AAV-2) has been determined to 3.0Ĺ resolution (Xie et al., 2002). AAV-2 crystallized in space group P1, with unit-cell parameters a = 249.7, b = 249.7, c = 644.8 Å, α = 90.0, β = 101.2, γ = 120.0. The crystals contained three full virus particles in the asymmetric unit, allowing 180-fold non-crystallographic symmetry averaging. The particle orientations were determined using the self-rotation function and found to have similar but resolvablv different orientations. Approximate alignment of icosahedral and interparticle threefold screw symmetry led to a native Patterson that was interpretable in terms of approximate particle positions. Accurate positions required a Patterson correlation search that was constrained to be consistent with non-crystallographic threefold projection symmetry evident in the diffraction intensities. Initial phases to 15.0 Å resolution were calculated by molecular replacement using the known structure of a distantly related homolog (23% sequence identity). Real-space averaging was performed and phases were extended from 15.0 to 3.0 Å. An atomic model was fitted and refined using a simulated-annealing real-space procedure.

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