In Situ Structural Analysis of Leptospira spp. by Electron Cryotomography.

Methods Mol Biol

Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.

Published: March 2021

Spirochetes such as Treponema, Borrelia, and Leptospira species can rotate their bodies to swim in liquid environments by rotating periplasmic flagella or endoflagella, which are present inside the cell. Electron cryotomography (ECT) is an imaging technique that directly provides three-dimensional (3D) structures of cells and molecular complexes in their cellular environment at nanometer resolution. Here, I present a general protocol of ECT that covers the sample preparation, data collection, tilt series alignment, and tomographic reconstruction for visualization of intact periplasmic flagella in Leptospira spp. This protocol is capable of determining protein structures at resolutions high enough to visualize their individual domains and secondary structures in their cellular environment.

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-0716-0459-5_12DOI Listing

Publication Analysis

Top Keywords

leptospira spp
8
electron cryotomography
8
periplasmic flagella
8
cellular environment
8
situ structural
4
structural analysis
4
analysis leptospira
4
spp electron
4
cryotomography spirochetes
4
spirochetes treponema
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!