Engineered Mutations Change the Structure and Stability of a Virus-Like Particle

The single-coat protein (CP) of bacteriophage Qβ self-assembles into T = 3 icosahedral virus-like particles (VLPs), of interest for a wide range of applications. These VLPs are very stable, but identification of the specific molecular determinants of this stability is lacking. To investigate these d...

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Bibliographic Details
Published in:Biomacromolecules 2012-08, Vol.13 (8), p.2339-2348
Main Authors: Fiedler, Jason D, Higginson, Cody, Hovlid, Marisa L, Kislukhin, Alexander A, Castillejos, Alexandra, Manzenrieder, Florian, Campbell, Melody G, Voss, Neil R, Potter, Clinton S, Carragher, Bridget, Finn, M.G
Format: Article
Language:English
Subjects:
Allolevivirus - genetics
Amino Acid Motifs
Amino Acid Substitution
Biological and medical sciences
Biotechnology
Capsid - chemistry
Capsid - ultrastructure
Capsid Proteins - biosynthesis
Capsid Proteins - chemistry
Capsid Proteins - genetics
Escherichia coli
Fundamental and applied biological sciences. Psychology
Hydrogen Bonding
Hydrophobic and Hydrophilic Interactions
Inverted Repeat Sequences
Light
Models, Molecular
Point Mutation
Protein Engineering
Protein Interaction Domains and Motifs
Protein Multimerization
Protein Stability
Protein Structure, Quaternary
Protein Unfolding
RNA, Viral - chemistry
RNA, Viral - genetics
Scattering, Radiation
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Summary:The single-coat protein (CP) of bacteriophage Qβ self-assembles into T = 3 icosahedral virus-like particles (VLPs), of interest for a wide range of applications. These VLPs are very stable, but identification of the specific molecular determinants of this stability is lacking. To investigate these determinants along with manipulations that confer more capabilities to our VLP material, we manipulated the CP primary structure to test the importance of various putative stabilizing interactions. Optimization of a procedure to incorporate fused CP subunits allowed for good control over the average number of covalent dimers in each VLP. We confirmed that the disulfide linkages are the most important stabilizing elements for the capsid and that acidic conditions significantly enhance the resistance of VLPs to thermal degradation. Interdimer interactions were found to be less important for VLP assembly than intradimer interactions. Finally, a single point mutation in the CP resulted in a population of smaller VLPs in three distinct structural forms.
ISSN:1525-7797
1526-4602
DOI:10.1021/bm300590x