Receptor-mediated uptake of Clostridioides difficile Toxins A and B
Final Report Abstract
The aim of the funded project was to reveal a possible synergy or hierarchy of the two known receptor binding regions of TcdB. Similarly, several interaction partners for TcdA are described in the literature, representing either a glycoprotein (Gp96) or different carbohydrate structures (sulfated glycosaminoglycans; Gal-Gal-GlcNac). Analogous to TcdB, (in)dependence of the receptor binding regions should also be investigated for this. Our results clearly demonstrate the complete independence of both receptor interactions for TcdB. Both toxin mutants with selective receptor specificity and CRISPR-Cas9-generated receptor-deficient cell lines confirm the completely independent function of both receptor binding regions in toxin uptake into the cell. In the context of elucidating an independent function of different receptor binding regions in TcdA, we were able to put the importance of sulfated glycosaminoglycans (sGAG) into perspective. The use of three different sGAG- deficient cell lines (HeLa, HT29, CHO) revealed that the uptake of TcdA was not significantly affected by the presence or absence of sGAG. The absence of the CROP domain (Gal-Gal- GlcNac binding) of TcdA or studies on cell lines showing decreased binding of the CROP domain reveal that TcdA from the reference strain VPI10463 and the hypervirulent strain R20291 have different affinity for the CROP-independent receptor. Even without knowing other specific receptors for TcdA, we can postulate the two-receptor model for TcdA as well. Here, both receptor interactions are independent and, as for TcdB, there appear to be different receptor affinities or even specificities for different TcdA variants.
Publications
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Evaluation of chondroitin sulfate proteoglycan-4 and frizzled-1,2,7 for cytopathic and cytotoxic effects of TcdB from historic and hypervirulent Clostridioides difficile strains. Naunyn-Schmiedeberg´s Archieve of Pharmacology, supplemental issue 1, 393:83 Abstracts der 5. German Pharm/Tox-Summit 2020 der Fachgesellschaft DGPT. DFG-Förderung GE 1017/6-1, 1. Jahr der Förderperiode
Henkel D.; Tatge H.; Schoettelndreier D.; Tao L.; Dong M. & Gerhard R.
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Receptor Binding Domains of TcdB from Clostridioides difficile for Chondroitin Sulfate Proteoglycan-4 and Frizzled Proteins Are Functionally Independent and Additive. Toxins, 12(12), 736.
Henkel, Daniel; Tatge, Helma; Schöttelndreier, Dennis; Tao, Liang; Dong, Min & Gerhard, Ralf
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Structural basis for CSPG4 as a receptor for TcdB and a therapeutic target in Clostridioides difficile infection. Nature Communications, 12(1).
Chen, Peng; Zeng, Ji; Liu, Zheng; Thaker, Hatim; Wang, Siyu; Tian, Songhai; Zhang, Jie; Tao, Liang; Gutierrez, Craig B.; Xing, Li; Gerhard, Ralf; Huang, Lan; Dong, Min & Jin, Rongsheng
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Identification of TFPI as a receptor reveals recombination-driven receptor switching in Clostridioides difficile toxin B variants. Nature Communications, 13(1).
Tian, Songhai; Xiong, Xiaozhe; Zeng, Ji; Wang, Siyu; Tremblay, Benjamin Jean-Marie; Chen, Peng; Chen, Baohua; Liu, Min; Chen, Pengsheng; Sheng, Kuanwei; Zeve, Daniel; Qi, Wanshu; Breault, David T.; Rodríguez, César; Gerhard, Ralf; Jin, Rongsheng; Doxey, Andrew C. & Dong, Min
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Impairment of lysosomal function by Clostridioides difficile TcdB. Molecular Microbiology, 117(2), 493-507.
Klepka, Carmen; Sandmann, Moritz; Tatge, Helma; Mangan, Matthew; Arens, Annabel; Henkel, Daniel & Gerhard, Ralf
