Nanoscale Visualization of Bacterial Microcompartments Using Atomic Force Microscopy
Bacterial microcompartments (BMCs) are polyhedral protein organelles in many prokaryotes, playing significant roles in metabolic enhancement. Due to their self-assembly and modularity nature, BMCs have gained increased interest in recent years, with the intent of constructing new nanobioreactors and scaffolding to promote cellular metabolisms and molecule delivery. In this chapter, we describe the technique of atomic force microscopy (AFM) as a method to study the self-assembly dynamics and physical properties of BMCs. We focus on the sample preparation, the measurement procedure, and the data analysis for high-speed AFM imaging and nanoindentation-based spectroscopy, which were used to determine the assembly dynamics of BMC shell proteins and the nanomechanics of intact BMC structures, respectively. The described methods could be applied to the study of other types of self-assembling biological organelles.
Key wordsAtomic force microscopy High-speed AFM Force spectroscopy Nanoindentation Bacterial microcompartment Carboxysome Self-assembly Nanomechanics
This work was supported by a Royal Society University Research Fellowship (UF120411), a Royal Society Research grant for University Research Fellowship (RG130442), a Royal Society Challenge grant (CH160004), a Biotechnology and Biological Sciences Research Council grant (BB/R003890/1), and a Biotechnology and Biological Sciences Research Council grant (BB/M024202/1). We acknowledge the Liverpool Centre for Cell Imaging for technical assistance and access to confocal/TIRF microscopes (Biotechnology and Biological Sciences Research Council, BB/M012441/1).
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