Production, purification and preliminary X-ray crystallographic studies of adeno-associated virus serotype 9.
Adeno-associated virus (AAV) serotype 9, which is under development for gene-delivery applications, shows significantly enhanced capsid-associated transduction efficiency in muscle compared with other AAV serotypes. With the aim of characterizing the structural determinants of this property, the purification, crystallization and preliminary X-ray crystallographic analyses of the AAV9 viral capsid are reported. The crystals diffracted X-rays to 2.8 A resolution using synchrotron radiation and belonged to the trigonal space group P3(2), with unit-cell parameters a = b = 251.0, c = 640.0 A. There are three complete viral capsids in the crystal unit cell. The orientation and position of the asymmetric unit capsid have been determined by molecular-replacement methods and structure determination is in progress.
Mitchell
Matthew
M
Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
Nam
Hyun-Joo
HJ
Carter
Adam
A
McCall
Angela
A
Rence
Chelsea
C
Bennett
Antonette
A
Gurda
Brittney
B
McKenna
Robert
R
Porter
Mark
M
Sakai
Yoshihisa
Y
Byrne
Barry J
BJ
Muzyczka
Nicholas
N
Aslanidi
George
G
Zolotukhin
Sergei
S
Agbandje-McKenna
Mavis
M
D002213:Capsid / Q000737:chemistry / Q000648:ultrastructure; D003460:Crystallization; D018360:Crystallography, X-Ray; D000229:Dependovirus / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification* / Q000648:ultrastructure; D012703:Serotyping