Structure of the cell-binding component of the Clostridium difficile binary toxin reveals a di-heptamer macromolecular assembly.
Xingjian XuRaquel Godoy-RuizKaylin A AdipietroChristopher PeraltaDanya Ben-HailKristen M VarneyMary E CookBraden M RothPaul T WilderThomas ClevelandAlexander GrishaevHeather M NeuSarah L J MichelWenbo YuDorothy BeckettRichard R RustandiCatherine LancasterJohn W LoughneyAdam KristopeitSianny ChristantiJessica W OlsonAlexander D MacKerellAmedee des GeorgesEdwin PozharskiDavid J WeberPublished in: Proceedings of the National Academy of Sciences of the United States of America (2020)
Targeting Clostridium difficile infection is challenging because treatment options are limited, and high recurrence rates are common. One reason for this is that hypervirulent C. difficile strains often have a binary toxin termed the C. difficile toxin, in addition to the enterotoxins TsdA and TsdB. The C. difficile toxin has an enzymatic component, termed CDTa, and a pore-forming or delivery subunit termed CDTb. CDTb was characterized here using a combination of single-particle cryoelectron microscopy, X-ray crystallography, NMR, and other biophysical methods. In the absence of CDTa, 2 di-heptamer structures for activated CDTb (1.0 MDa) were solved at atomic resolution, including a symmetric (SymCDTb; 3.14 Å) and an asymmetric form (AsymCDTb; 2.84 Å). Roles played by 2 receptor-binding domains of activated CDTb were of particular interest since the receptor-binding domain 1 lacks sequence homology to any other known toxin, and the receptor-binding domain 2 is completely absent in other well-studied heptameric toxins (i.e., anthrax). For AsymCDTb, a Ca2+ binding site was discovered in the first receptor-binding domain that is important for its stability, and the second receptor-binding domain was found to be critical for host cell toxicity and the di-heptamer fold for both forms of activated CDTb. Together, these studies represent a starting point for developing structure-based drug-design strategies to target the most severe strains of C. difficile.
Keyphrases
- escherichia coli
- clostridium difficile
- binding protein
- high resolution
- biofilm formation
- single cell
- cell therapy
- magnetic resonance
- magnetic resonance imaging
- stem cells
- early onset
- computed tomography
- klebsiella pneumoniae
- emergency department
- cell proliferation
- optical coherence tomography
- cancer therapy
- transcription factor
- ionic liquid
- mass spectrometry
- hydrogen peroxide
- multidrug resistant
- protein kinase
- solid state
- amino acid
- case control