Protocol for imaging proteins associated with Legionella-containing vacuoles in host cells.

Abstract

The intracellular bacterial pathogen Legionella pneumophila exploits host cellular systems using approximately 300 effector proteins to establish a replicative niche known as the Legionella-containing vacuole (LCV). During infection, both host and bacterial proteins interactively function on the LCVs. Here, we describe a detailed step-by-step protocol to visualize proteins associated with LCVs in host cells. This protocol can aid in analyzing whether a protein of interest influences the subcellular localization of LCV-associated proteins during infection. For complete details on the use and execution of this protocol, please refer to Kitao et al. (2020).

• Authors: Tomoe Kitao, Kohei Arasaki, Hiroki Nagai, Tomoko Kubori
• Journal: STAR protocols (STAR Protoc) Vol. 2 Issue 2 Pg. 100410 (06 18 2021) ISSN: 2666-1667 [Electronic] United States
• PMID: 33870219 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2021 The Author(s).

Chemical References

• Bacterial Proteins
• Green Fluorescent Proteins

Topics

• Bacterial Proteins (analysis, chemistry)
• Bacteriological Techniques (methods)
• Cell Culture Techniques
• Fluorescent Antibody Technique (methods)
• Green Fluorescent Proteins (genetics, metabolism)
• HEK293 Cells
• Host-Pathogen Interactions (physiology)
• Humans
• Legionella pneumophila (chemistry)
• Plasmids (genetics, metabolism)
• Transfection
• Vacuoles (chemistry, microbiology)