Light-Controlled Secretion and Injection of Proteins into Eukaryotic Cells by Optogenetic Control of the Bacterial Type III Secretion System

Florian Lindner; Sebastian Grossmann; Andreas Diepold

doi:10.1007/978-1-0716-4047-0_9

Light-Controlled Secretion and Injection of Proteins into Eukaryotic Cells by Optogenetic Control of the Bacterial Type III Secretion System

Methods Mol Biol. 2025:2840:115-131. doi: 10.1007/978-1-0716-4047-0_9.

Authors

Florian Lindner¹, Sebastian Grossmann¹, Andreas Diepold^{2

3}

Affiliations

¹ Department of Ecophysiology, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
² Department of Ecophysiology, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany. [email protected].
³ Center for Synthetic Microbiology (SYNMIKRO), Marburg, Germany. [email protected].

PMID: 39724348
DOI: 10.1007/978-1-0716-4047-0_9

Abstract

Gram-negative bacteria can use the type III secretion system (T3SS) to inject effector proteins into eukaryotic target cells. In this chapter, we describe the application of a light-controlled T3SS, based on the targeted sequestration of an essential dynamic T3SS component with the help of optogenetic interaction switches. This method enables to control the secretion or injection into eukaryotic cells for a wide range of protein cargos with high temporal and spatial precision.

Keywords: Bacterial secretion systems; Cellular control; Injectisome; LITESEC-T3SS; LOVTRAP; Membrane transport; Optogenetic interaction switches; Protein translocation; iLID.

MeSH terms

Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Eukaryotic Cells / metabolism
Humans
Light*
Optogenetics* / methods
Protein Transport
Type III Secretion Systems* / genetics
Type III Secretion Systems* / metabolism

Substances

Type III Secretion Systems
Bacterial Proteins