CryoEM of bacterial secretion systems: A primer for microbiologists. Issue 3 (5th December 2020)

• Record Type: Journal Article
• Authors: Umrekar, Trishant R.; Cohen, Eli; Drobnič, Tina; Gonzalez‐Rodriguez, Nayim; Beeby, Morgan
• Other Names: Blokesch Melanie guestEditor.; Palmer Tracy guestEditor.
• Is Part Of: Molecular microbiology. Volume 115:Issue 3(2021)
• Page Start: 366
• Page End: 382
• Publication Date: 2020-12-05
• Subjects: bacterial secretion systems -- cryoEM -- electron cryotomography -- single particle analysis; Molecular microbiology -- Periodicals; 572.829
• DOI: 10.1111/mmi.14637
• Languages: English
• ISSNs: 0950-382X

Abstract

Abstract: "CryoEM" has come of age, enabling considerable structural insights into many facets of molecular biology. Here, we present a primer for microbiologists to understand the capabilities and limitations of two complementary cryoEM techniques for studying bacterial secretion systems. The first, single particle analysis, determines the structures of purified protein complexes to resolutions sufficient for molecular modeling, while the second, electron cryotomography and subtomogram averaging, tends to determine more modest resolution structures of protein complexes in intact cells. We illustrate these abilities with examples of insights provided into how secretion systems work by cryoEM, with a focus on type III secretion systems. Abstract : Main stages of the cryoEM workflow: samples (either purified molecules, or intact cells) are flash frozen, imaged in the electron microscope, and the resulting 2‐D projection images used to reconstruct a 3‐D structure of the sample.