Purification
and Characterization of a Stable, Membrane-Associated
Peptidoglycan Responsive Adenylate Cyclase LRR Domain from Human Commensal Candida albicans
posted on 2022-12-20, 08:29authored byGeneva
Maddison Crump, Sharon Rozovsky, Catherine L. Grimes
The evolutionarily conserved leucine rich repeat (LRR)
protein
domain is a unique structural motif found in many viral, bacterial,
archaeal, and eukaryotic proteins. The LRR domain serves many roles,
including being a signaling domain and a pathogen recognition receptor.
In the human innate immune system, it serves an essential role by
recognizing fragments of bacterial cell walls. Interestingly, the
human fungal pathogen Candida albicans also uses
an LRR domain-containing protein, Cyrp1, to sense bacterial cell wall
fragments. However, the dynamics of signaling and detection of bacterial
peptidoglycan fragments by the LRR of Cyr1p remains poorly characterized.
Here we develop optimal recombinant expression workflows and provide
characterization of the entire region of the LRR domain of Cyr1p as
a peripheral membrane protein. Using a newly designed peptidoglycan
enrichment bead assay, we demonstrate that this domain can bind bacterial
peptidoglycan fragments under native conditions. The new membrane-associated
Cyr1p–LRR construct sets the stage for the development of antifungal
agents via high-throughput campaigns to inhibit cell wall–Cyr1p
interactions.