Purification and Cryo-Electron Microscopy Analysis of Bacterial Appendages.

ABSTRACT: A number of extracellular helical protein polymers are crucial for supporting bacterial motility. The bacterial flagellum is a polymeric appendage used to support cellular motility. Historically, structural studies of flagellar and other filaments were limited to those present as or locked into straightened states. Here, we present a robust workflow that produces biologically relevant high-resolution cryo-electron microscopy (cryo-EM) structures of bacterial flagellar filaments. We highlight how a simple purification method, centered around several centrifugation steps, exploits the process of filament ejection in Caulobacter crescentus and results in isolated filaments amenable to transmission electron microscopy (TEM) studies. The quality of the sample is validated by SDS-PAGE and

SUBMITTER: Sanchez JC 

PROVIDER: S-EPMC11292135 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature