Structural insight into the unexploited allosteric binding site of fructose 1, 6-bisphosphate aldolase from C. albicans with α-lipoic acid.

The rising incidence of drug-resistant fungal infections underscores the urgent need for innovative therapeutic strategies. However, developing selective treatments remains a formidable challenge due to the similarities between humans and fungal cells. Class II fructose 1,6-bisphosphate aldolase (FBA) represents an attractive pharmacological target for the development of antifungal agents due to its crucial role in microbial survival and its absence in human. In this work, we identified α-lipoic acid (ALA), a naturally occurring compound, as a novel inhibitor of C. albicans FBA (CaFBA). The co-crystallography, enzyme inhibition assays, and site-directed mutagenesis revealed that ALA acts as a non-covalent inhibitor, binding to an unexploited allosteric site on CaFBA, distinct from the previously reported substrate-binding pocket or C292 covalent binding site. Notably, ALA selectively inhibits CaFBA, likely due to the non-conservation of the allosteric binding site, particularly S268, across species. The synergistic inhibition of C. albicans by ALA and amphotericin B highlights its therapeutic potential as part of a combined antifungal strategy. In summary, this study provides a structural basis for the design and optimization of novel CaFBA inhibitors, enhancing our understanding of FBA's role in fungal growth and establishing a foundation for developing effective antifungal therapeutics against C. albicans.