Bacteria employ a range of tools and mechanisms to infect and damage host cells, evade immune systems, and protect against toxic compounds like heavy metal ions and antimicrobials. The expulsion of the latter is critical to antimicrobial resistance strategies, and is commonly facilitated by membrane-spanning efflux pumps. In Gram-negative bacteria, these efflux pumps are often tripartite complexes composed of an inner membrane complex, a periplasmic adaptor protein, and an outer membrane factor such as the TolC protein-- a crucial player in the cell’s ability to adapt to challenging environments, most notably during the first stages of resistance to clinical drugs. While the TolC is found exclusively in Gram-negative bacteria, members of the TolC-like protein family were recently discovered in Gram-positives like Clostridium difficile which lack an outer membrane. Instead, these TolC-like proteins reside within the bacterial surface layer, a protein coat involved in host cell adherence and antimicrobial protection, which raises the questions: do these proteins serve the same purpose as their Gram-negative counterparts? And how do their structures differ?