Isolation, Characterization, and Aggregation of a Structured Bacterial Matrix Precursor

Abstract

Biofilms are surface-associated groups of microbial cells that are embedded in an extracellular matrix (ECM). The ECM is a network of biopolymers, mainly polysaccharides, proteins, and nucleic acids. ECM proteins serve a variety of structural roles and often form amyloid-like fibers. Despite the extensive study of the formation of amyloid fibers from their constituent subunits in humans, much less is known about the assembly of bacterial functional amyloid-like precursors into fibers. Using dynamic light scattering, atomic force microscopy, circular dichroism, and infrared spectroscopy, we show that our unique purification method of a Bacillus subtilis major matrix protein component results in stable oligomers that retain their native α-helical structure. The stability of these oligomers enabled us to control the external conditions that triggered their aggregation. In particular, we show that stretched fibers are formed on a hydrophobic surface, whereas plaque-like aggregates are formed in solution under acidic pH conditions. TasA is also shown to change conformation upon aggregation and gain some β-sheet structure. Our studies of the aggregation of a bacterial matrix protein from its subunits shed new light on assembly processes of the ECM within bacterial biofilms. Background: TasA is an extracellular matrix protein that makes amyloid-like fibers in Bacillus subtilis biofilms. Results: An isolated TasA matrix precursor self-assembled in vitro into fibers on hydrophobic surfaces and in acidic solutions. Conclusion: TasA is purified as stable, structured oligomers that aggregate in response to simple physical external cues. Significance: TasA aggregation principles can be used to design new anti-biofilm drugs and surfaces.