In Vitro Flagellar Type III Protein Transport Assay Using Inverted Membrane Vesicles

Katsumi Imada; Hiroyuki Terashima

doi:10.1007/978-1-0716-3060-0_2

In Vitro Flagellar Type III Protein Transport Assay Using Inverted Membrane Vesicles

Methods Mol Biol. 2023:2646:17-26. doi: 10.1007/978-1-0716-3060-0_2.

Authors

Katsumi Imada¹, Hiroyuki Terashima²

Affiliations

¹ Department of Macromolecular Science, Graduate School of Science, Osaka University, Toyonaka, Osaka, Japan. kimada@chem.sci.osaka-u.ac.jp.
² Department of Bacteriology, Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.

PMID: 36842102
DOI: 10.1007/978-1-0716-3060-0_2

Abstract

The flagellar axial proteins are transported across the cytoplasmic membrane into the central channel of the growing flagellum via the flagellar protein export apparatus, a member of the type III secretion system (T3SS). To reveal the molecular mechanism of protein transport by the T3SS, accurate measurement of protein transport under various conditions is essential. In this chapter, we describe an in vitro method for flagellar protein transport assay using inverted membrane vesicles (IMVs) prepared from Salmonella cells. This method can easily and precisely control the condition around the T3SS and be applied to other T3SSs.

Keywords: Inverted membrane vesicle; Protein transport assay; T3SS.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins* / metabolism
Biological Transport
Cell Membrane / metabolism
Flagella* / metabolism
Membranes / metabolism
Protein Transport

Substances

Bacterial Proteins