Engineering enhanced protein disaggregases for neurodegenerative disease.
Protein misfolding and aggregation underpin several fatal neurodegenerative diseases, including Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and frontotemporal dementia (FTD). There are no treatments that directly antagonize the protein-misfolding events that cause these disorders. Agents that reverse protein misfolding and restore proteins to native form and function could simultaneously eliminate any deleterious loss-of-function or toxic gain-of-function caused by misfolded conformers. Moreover, a disruptive technology of this nature would eliminate self-templating conformers that spread pathology and catalyze formation of toxic, soluble oligomers. Here, we highlight our efforts to engineer Hsp104, a protein disaggregase from yeast, to more effectively disaggregate misfolded proteins connected with PD, ALS, and FTD. Remarkably subtle modifications of Hsp104 primary sequence yielded large gains in protective activity against deleterious α-synuclein, TDP-43, FUS, and TAF15 misfolding. Unusually, in many cases loss of amino acid identity at select positions in Hsp104 rather than specific mutation conferred a robust therapeutic gain-of-function. Nevertheless, the misfolding and toxicity of EWSR1, an RNA-binding protein with a prion-like domain linked to ALS and FTD, could not be buffered by potentiated Hsp104 variants, indicating that further amelioration of disaggregase activity or sharpening of substrate specificity is warranted. We suggest that neuroprotection is achievable for diverse neurodegenerative conditions via surprisingly subtle structural modifications of existing chaperones.
a Department of Biochemistry and Biophysics ; Perelman School of Medicine at the University of Pennsylvania ; Philadelphia , PA USA.
Meredith E
ME
Jackrel
James
J
Shorter
D000682:Amyloid / Q000378:metabolism*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D019636:Neurodegenerative Diseases / Q000378:metabolism*; D015202:Protein Engineering; D017510:Protein Folding; D034802:RNA-Binding Protein EWS; D016601:RNA-Binding Proteins / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
D000682:Amyloid; D002148:Calmodulin-Binding Proteins; C500986:EWSR1 protein, human; D006360:Heat-Shock Proteins; D034802:RNA-Binding Protein EWS; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; C070972:HsP104 protein, S cerevisiae
AD, Alzheimer's disease; ALS, amyotrophic lateral sclerosis; EWSR1; FTD, frontotemporal dementia; FUS; HD, Huntington's disease; Hsp104; MD, middle domain; NBD, nucleotide-binding domain; PD, Parkinson's disease; TAF15; TDP-43; WT, wild-type; amyloid; prion; protein disaggregation; protein engineering; α-syn, α-synuclein; α-synuclein