Publication: Carbene Footprinting maps GSDMD-Caspase binding site

Mar 18, 2024

“Mapping the Binding Interactions between Human Gasdermin D and Human Caspase-1 Using Carbene Footprinting”

Work from Professor Neil Oldham’s research group on Carbene Footprinting has recently been published in JACS Au, mapping the binding sites of Gasdermin-D, Caspase-1 and a small molecule inhibitor.

The research, performed in collaboration with scientists from OMass Therapeutics, focuses on the interactions between wild-type Gasdermin and Caspase. Gasdermin-D, upon activation by Caspase, is a pore forming protein. Gasdermin-D activation therefore is one of the key initiators of pyroptosis, a contributor towards cancers and inflammatory diseases. Inhibition of this pathway is seen as a key therapeutic strategy.

Carbene footprinting was used to show the binding between Gasdermin-D and Caspase. The technique uses a laser to generate a highly reactive carbene species from a diazirine functional group, which subsequently binds to the accessible amino acids on the proteins surface. When the protein is bound to its partner, this binding site becomes inaccessible to the carbene species, and therefore cannot react. By monitoring the differences in carbene distribution across the protein surface, Carbene Footprinting is used to map the binding sites of the protein with its partner. You can read more about Carbene Footprinting here.

VRT-043198, which modifies the Cystine at position 285 and therefore inhibits Caspase’s activation of Gasdermin-D. Carbene Footprinting was used to map the non-covalent binding of VRT-043198 onto a mutated form of Caspase (C285A). The data confirms that despite the mutation meaning the compound can no longer covalently modify the Cystine, non-covalent binding between the mutant caspase and VRT-043198 can still be observed in the same binding site.

You can read the paper in full by clicking here.

You can find a full list of our papers by clicking here. 

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