Conversion of the OmpF Porin into a Device to Gather Amyloids on the E. coli Outer Membrane

Area of Science:

• Biochemistry
• Microbiology
• Materials Science

Background:

• Protein amyloids are widespread and pose risks due to infectivity and toxicity.
• Current concerns involve their role in gut microbiota and soil environments.
• Understanding amyloid interactions is crucial for developing mitigation strategies.

Purpose of the Study:

• To engineer a novel bacterial system for trapping protein amyloids.
• To explore the potential of modified outer membrane porins for amyloid detection and clearance.
• To investigate the mechanism of amyloid-peptide recognition by engineered bacteria.

Main Methods:

• Insertion of an amyloidogenic peptide sequence from RepA-WH1 into the L5 loop of Escherichia coli OmpF.
• Expression of the engineered OmpF in E. coli.
• Demonstration of bacterial cell trapping of free amyloidogenic peptides.
• Utilizing immobilized prion-like proteins to capture engineered bacteria.
• Testing the effect of polyphenolic inhibitors on peptide recognition.

Main Results:

• Engineered E. coli successfully trapped extracellular amyloidogenic peptides on their surface.
• The modified OmpF facilitated homotypic interactions characteristic of amyloid assembly.
• Immobilized prion-like proteins could capture bacteria displaying the engineered OmpF.
• Polyphenolic molecules interfered with the recognition process, validating the amyloid-like interaction.

Conclusions:

• Synthetic porins can be engineered to interact with specific protein sequences.
• This approach offers a platform for developing biosensors and clearance devices for pathogenic amyloids.
• The study highlights a novel strategy for managing amyloid-related threats in various environments.