8A9U

Full AAV3B-VP1KO virion

8A9U の概要

• エントリーDOI: 10.2210/pdb8a9u/pdb
• EMDBエントリー: 15286
• 分子名称: Capsid protein VP1 (1 entity in total)
• 機能のキーワード: adeno-associated virus, aav, aav3b, aav serotype 3b, full, vp1ko, virus
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 66728.28

構造登録者

Arriaga, I.,Abrescia, N.G.A. (登録日: 2022-06-29, 公開日: 2022-09-21, 最終更新日: 2024-11-13)

主引用文献

Arriaga, I.,Navarro, A.,Etxabe, A.,Trigueros, C.,Samulski, R.J.,Moullier, P.,Francois, A.,Abrescia, N.G.A.
Cellular and Structural Characterization of VP1 and VP2 Knockout Mutants of AAV3B Serotype and Implications for AAV Manufacturing.
Hum Gene Ther, 33:1142-1156, 2022

PubMed Abstract: AAV virion biology is still lacking a complete understanding of the role that the various structural subunits (VP1, 2, and 3) play in virus assembly, infectivity, and therapeutic delivery for clinical indications. In this study, we focus on the less studied adeno-associated virus AAV3B and generate a collection of AAV plasmid substrates that assemble virion particles deficient specifically in VP1, VP2, or VP1 and 2 structural subunits. Using a collection of biological and structural assays, we observed that virions devoid of VP1, VP2, or VP1 and 2 efficiently assembled virion particles, indistinguishable by cryoelectron microscopy (cryo-EM) from that of wild type (WT), but unique in virion transduction (WT > VP2 > VP1 > VP1 and 2 mutants). We also observed that the missing structural subunit was mostly compensated by additional VP3 protomers in the formed virion particle. Using cryo-EM analysis, virions fell into three classes, namely full, empty, and partially filled, based on comparison of density values within the capsid. Further, we characterize virions described as "broken" or "disassembled" particles, and provide structural information that supports the particle dissolution occurring through the two-fold symmetry sites. Finally, we highlight the unique value of employing cryo-EM as an essential tool for release criteria with respect to AAV manufacturing.

PubMed: 36082996
DOI: 10.1089/hum.2022.119

実験手法

ELECTRON MICROSCOPY (3.1 Å)