Production, purification, crystallization and preliminary X-ray structural studies of adeno-associated virus serotype 5.
Adeno-associated virus serotype 5 (AAV5) is under development for gene-therapy applications for the treatment of cystic fibrosis. To elucidate the structural features of AAV5 that control its enhanced transduction of the apical surface of airway epithelia compared with other AAV serotypes, X-ray crystallographic studies of the viral capsid have been initiated. The production, purification, crystallization and preliminary crystallographic analysis of empty AAV5 viral capsids are reported. The crystals diffract X-rays to beyond 3.2 A resolution using synchrotron radiation and belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 264.7, b = 447.9, c = 629.7 A. There is one complete T = 1 viral capsid per asymmetric unit. The orientation and position of the viral capsid in the asymmetric unit have been determined by rotation and translation functions, respectively, and the AAV5 structure determination is in progress.
DiMattia
Michael
M
Department of Biochemistry and Molecular Biology, McKnight Brain Institute, Center for Structural Biology, University of Florida, Gainesville, FL 32610, USA.
Govindasamy
Lakshmanan
L
Levy
Hazel C
HC
Gurda-Whitaker
Brittney
B
Kalina
Amy
A
Kohlbrenner
Erik
E
Chiorini
John A
JA
McKenna
Robert
R
Muzyczka
Nicholas
N
Zolotukhin
Sergei
S
Agbandje-McKenna
Mavis
M
D002213:Capsid / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D000229:Dependovirus / Q000648:ultrastructure*; D015316:Genetic Therapy; D008854:Microscopy, Electron; D046529:Microscopy, Electron, Transmission; D016366:Open Reading Frames; D017356:Synchrotrons; D014764:Viral Proteins / Q000737:chemistry
D014764:Viral Proteins