Multiple parameters beyond lipid binding affinity drive cytotoxicity of cholesterol-dependent cytolysins

Sucharit Ray, Roshan Thapa, Peter A. Keyel

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

The largest superfamily of bacterial virulence factors is pore-forming toxins (PFTs). PFTs are secreted by both pathogenic and non-pathogenic bacteria. PFTs sometimes kill or induce pro-pathogen signaling in mammalian cells, all primarily through plasma membrane perforation, though the parameters that determine these outcomes are unclear. Membrane binding, calcium influx, pore size, and membrane repair are factors that influence PFT cytotoxicity. To test the contribution of membrane binding to cytotoxicity and repair, we compared the closely related, similarly-sized PFTs Perfringolysin O (PFO) from Clostridium perfringens and Streptolysin O (SLO) from Streptococcus pyogenes. Cell death kinetics for PFO and SLO were different because PFO increased in cytotoxicity over time. We introduced known L3 loop mutations that swap binding affinity between toxins and measured hemolytic activity, nucleated cell death kinetics and membrane repair using viability assays, and live cell imaging. Altered hemolytic activity was directly proportional to toxin binding affinity. In contrast, L3 loop alterations reduced nucleated cell death, and they had limited effects on cytotoxicity kinetics and membrane repair. This suggests other toxin structural features, like oligomerization, drives these parameters. Overall, these findings suggest that repair mechanisms and toxin oligomerization add constraints beyond membrane binding on toxin evolution and activity against nucleated cells.

Original languageEnglish
Article number1
JournalToxins
Volume11
Issue number1
DOIs
StatePublished - Jan 2019

Keywords

  • Clostridium perfringens
  • Intrinsic repair
  • Membrane repair
  • Microvesicle shedding
  • Perfringolysin
  • Pore-forming toxin
  • Streptococcus pyogenes
  • Streptolysin

Fingerprint

Dive into the research topics of 'Multiple parameters beyond lipid binding affinity drive cytotoxicity of cholesterol-dependent cytolysins'. Together they form a unique fingerprint.

Cite this