Monitoring protien cage nanopaticle morphology for applications in medicines and materials

Abstract

Protein cage nanoparticles are naturally occurring proteins found in all domains of life. The breadth of structural knowledge and the ability to modify protein cage nanoparticles both chemically and genetically set them apart for use as platforms for biomedical templates and materials synthesis. The work described herein focuses on the use of protein cage nanoparticles as a protective agent from a suite of viral pathogens. Protein cage nanoparticles exist in many different morphological forms both within a specific particle and between particles. It is essential to characterize these different states in order to engineer a protein cage nano particle for biomedical and materials synthesis. Described here is an expanded protocol for determining the morphological state with the bacteriophage P22 capsid. Using multiple techniques including multi angle light scattering, analytical ultra centrifugation, agarose gel electrophoresis and transmission electron microscopy these states are described and characterized. P22 exits in four different morphological states: the procapsid, empty shell, expanded shell and so-called "wiffleball". Also characterized in the work is the small heat shock protein from Methanococcus jannaschii, which exists in two morphological states. One of the states being the assembled 12 nm cage structure and the other state being a disassembled cage structure that is most commonly described at elevated temperatures. The characterization of these structures can aid in the understanding the mechanism of formation for the immunological phenomena induced bronchial associated lymphoid tissue.