Detection of intermediates and kinetic control during assembly of bacteriophage P22 procapsid

JOURNAL OF MOLECULAR BIOLOGY

卷:381

Article

Tuma, Roman^1,2,3  Tsuruta, Hiro⁴  French, Kenneth H.²  Prevelige, Peter E.² 

[1] Univ Leeds, Astbury Ctr Struct Mol Biol, Leeds, W Yorkshire, England

[2] Univ Alabama, Dept Microbiol, Birmingham, AL 35294 USA

[3] Univ Helsinki, Inst Biotechnol, FIN-00014 Helsinki, Finland

[4] Stanford Univ, Stanford Linear Accelerator Ctr, Stanford Synchrotron Radiat Lab, Menlo Pk, CA 94025 USA

关键词: self-assembly;    virus;    X-ray;    kinetics;    structure;   

DOI  :  10.1016/j.jmb.2008.06.020

来源: Elsevier

【 摘 要 】

Bacteriophage P22 serves as a model for the assembly and maturation of other icosahedral double-stranded DNA viruses. P22 coat and scaffolding proteins assemble in vitro into an icosahedral procapsid, which then expands during DNA packaging (maturation). Efficient in vitro assembly makes this system suitable for design and production of monodisperse spherical nanoparticles (diameter approximate to 50 nm). In this work, we explore the possibility of controlling the outcome of assembly by scaffolding protein engineering. The scaffolding protein exists in monomer-dimer-tetramer equilibrium. We address the role of monomers and dimers in assembly by using three different scaffolding proteins with altered monomer-dimer equilibrium (weak dimer, covalent dimer, monomer). The progress and outcome of assembly was monitored by time-resolved X-ray scattering, which allowed us to distinguish between closed shells and incomplete assembly intermediates. Binding of scaffolding monomer activates the coat protein for assembly. Excess dimeric scaffolding protein resulted in rapid nucleation and kinetic trapping yielding incomplete shells. Addition of monomeric wild-type scaffold with excess coat protein completed these metastable shells. Thus, the monomeric scaffolding protein plays an essential role in the elongation phase by activating the coat and effectively lowering its critical concentration for assembly. (c) 2008 Elsevier Ltd. All rights reserved.

【 授权许可】

Free