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Mechanism of germination inhibition of Clostridioides difficile spores by an aniline substituted cholate derivative (CaPA)

The Journal of Antibiotics volume 76, pages 335–345 (2023)Cite this article

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Abstract

Clostridioides difficile infection (CDI) is the major identifiable cause of antibiotic-associated diarrhea and has been declared an urgent threat by the CDC. C. difficile forms dormant and resistant spores that serve as infectious vehicles for CDI. To cause disease, C. difficile spores recognize taurocholate and glycine to trigger the germination process. In contrast to other sporulating bacteria, C. difficile spores are postulated to use a protease complex, CspABC, to recognize its germinants. Since spore germination is required for infection, we have developed anti-germination approaches for CDI prophylaxis. Previously, the bile salt analog CaPA (an aniline-substituted cholic acid) was shown to block spore germination and protect rodents from CDI caused by multiple C. difficile strains and isolates. In this study, we found that CaPA is an alternative substrate inhibitor of C. difficile spore germination. By competing with taurocholate for binding, CaPA delays C. difficile spore germination and reduces spore viability, thus diminishing the number of outgrowing vegetative bacteria. We hypothesize that the reduction of toxin-producing bacterial burden explains CaPA’s protective activity against murine CDI. Previous data combined with our results suggests that CaPA binds tightly to C. difficile spores in a CspC-dependent manner and irreversibly traps spores in an alternative, time-delayed, and low yield germination pathway. Our results are also consistent with kinetic data suggesting the existence of at least two distinct bile salt binding sites in C. difficile spores.

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Acknowledgements

This work was supported by the National Institute of Health [grant numbers R01-AI109139 and GM103440]. The authors thank Prof. Steve Firestine from Wayne State University for the synthesis of CaPA, Prof. Nigel Minton from University of Nottingham for providing clinical C. difficile isolates, and Prof. Aimee Shen from Tufts University for the training and materials for C. difficile knock-out system.

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  1. Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, NV, 89154, USA

    Christopher Yip, Jacqueline R. Phan & Ernesto Abel-Santos

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Correspondence to Ernesto Abel-Santos.

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Yip, C., Phan, J.R. & Abel-Santos, E. Mechanism of germination inhibition of Clostridioides difficile spores by an aniline substituted cholate derivative (CaPA). J Antibiot 76, 335–345 (2023). https://doi.org/10.1038/s41429-023-00612-3

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