Fluorescent Functionalization across Quaternary Structure in a Virus-like Particle

5 years ago

Fluorescent Functionalization across Quaternary Structure in a Virus-like Particle

Galo L. Mejia, Kyle W. Murray, Sheena D’Arcy, Duane D. Winkler, Raymond P. Welch, Gabriele Meloni, Candace E. Benjamin, Zhuo Chen, Jenifer S. Calvo, Stefanie D. Boyd, Jeremiah J. Gassensmith

Proteinaceous nanomaterials and, in particular, virus-like particles (VLPs) have emerged as robust and uniform platforms that are seeing wider use in biomedical research. However, there are a limited number of bioconjugation reactions for functionalizing the capsids, and very few of those involve functionalization across the supramolecular quaternary structure of protein assemblies. In this work, we exploit the recently described dibromomaleimide moiety as part of a bioconjugation strategy on VLP Qβ to break and rebridge the exposed and structurally important disulfides in good yields. Not only was the stability of the quaternary structure retained after the reaction, but the newly functionalized particles also became brightly fluorescent and could be tracked in vitro using a commercially available filter set. Consequently, we show that this highly efficient bioconjugation reaction not only introduces a new functional handle “between” the disulfides of VLPs without compromising their thermal stability but also can be used to create a fluorescent probe.

Publisher URL: http://dx.doi.org/10.1021/acs.bioconjchem.7b00305

DOI: 10.1021/acs.bioconjchem.7b00305