University of California: San Francisco
GPCRs! DMS! More coming soon 😉
Washington University in St. Louis:
Modeling PSM⍺1-4 amyloidogenesis and disrupting it using engineered protein disaggregases
Phenol Soluble Modulins (PSMs) are a key family of secreted proteins from S. aureus that hinder immune responses and contribute to biofilm stability via the formation of amyloid fibers. We developed a yeast system to study their aggregation and toxicity that allowed us to simultaneously screen engineered Hsp104 variants. Using this system, we were able to identify potentiated Hsp104 variants capable of disrupting PSM aggregates to promote their clearance. Importantly, these same Hsp104 disaggregases also act on preformed S. aureus biofilms to promote their disassembly. This suggests that Hsp104 disaggregases could be used a novel enzymatic method of biofilm disruption and clearance.
Functional Analysis of Hsp104 Pore Loops
Hsp104 is a hexameric AAA+ yeast disaggregase capable of solubilizing disordered aggregates and amyloid. Hsp104 couples ATP hydrolysis to polypeptide translocation through its
central channel, which is mediated by conserved tyrosine residues in nucleotide binding domain (NBD) 1 and NBD2. Here, we analyze the properties of a proposed third interaction mediated by tyrosine 650. Through a series of functional assays, we suggest that Y650 is not essential for Hsp104 function, but that its modulation may be useful for fine-tuning and engineering the properties of Hsp104.