Assembling the Bacillus subtilis Spore Coat Basement Layer on Spherical Supported Lipid Bilayers

Methods Mol Biol. 2024:2727:215-225. doi: 10.1007/978-1-0716-3491-2_17.

Abstract

Micro- and nanoparticles are often designed by mimicking naturally occurring structures. Bacterial spores are dormant cells elaborated by some Gram-positive bacteria during poor growth conditions to protect their genetic material from harsh environmental stresses. In Bacillus subtilis, this protection is, in part, conferred by a proteinaceous shell, the "coat", which is composed of ~80 different proteins. The basement layer of the coat contains two unusual proteins, which we have recently reconstituted around silica beads to generate synthetic spore-like particles termed "SSHELs". Here, we describe the protocol for generating SSHEL particles, and describe the procedure to covalently link molecules of interest (in this case an anti-HER2 affibody) to SSHEL surfaces. SSHELs therefore represent a versatile platform for the display of ligands or antigens for the site-specific delivery of cargo or vaccines.

Keywords: C. difficile; Cancer; Drug delivery; Nanoparticle; SpoIVA; SpoVM; Sporulation; Synthetic biology.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis* / metabolism
  • Bacterial Proteins / metabolism
  • Cell Wall / metabolism
  • Lipid Bilayers* / metabolism
  • Spores, Bacterial / genetics

Substances

  • Lipid Bilayers
  • Bacterial Proteins