Engineered mutations change the structure and stability of a virus-like particle |
| |
Authors: | Jason D Fiedler Cody Higginson Marisa L Hovlid Alexander A Kislukhin Alexandra Castillejos Florian Manzenrieder Melody G Campbell Neil R Voss Clinton S Potter Bridget Carragher M G Finn |
| |
Affiliation: | Department of Chemistry and ?The National Resource for Automated Molecular Microscopy, Department of Cell Biology, The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States. |
| |
Abstract: | 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. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|