A kinetic assessment of the C. elegans amyloid disaggregation activity enables uncoupling of disassembly and proteolysis.
Protein aggregation is a common feature of late onset neurodegenerative disorders, including Alzheimer's disease. In Alzheimer's disease, misassembly of the Abeta peptide is genetically linked to proteotoxicity associated with disease etiology. A reduction in Abeta proteotoxicity is accomplished, in part, by the previously reported Abeta disaggregation and proteolysis activities-under partial control of heat shock factor 1, a transcription factor regulating proteostasis in the cytosol and negatively regulated by insulin growth factor signaling. Herein, we report an improved in vitro assay to quantify recombinant fibrillar Abeta disaggregation kinetics accomplished by the exogenous application of C.elegans extracts. With this assay we demonstrate that the Abeta disaggregation and proteolysis activities of C.elegans are separable. The disaggregation activity found in C.elegans preparations is more heat resistant than the proteolytic activity. Abeta disaggregation in the absence of proteolysis was found to be a reversible process. Future discovery of the molecular basis of the disaggregation and proteolysis activities offers the promise of delaying the age-onset proteotoxicity that leads to neurodegeneration in a spectrum of maladies.
Bieschke
Jan
J
Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Cohen
Ehud
E
Murray
Amber
A
Dillin
Andrew
A
Kelly
Jeffery W
JW
D000544:Alzheimer Disease; D016229:Amyloid beta-Peptides / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000737:chemistry* / Q000201:enzymology; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry / Q000378:metabolism*; D004195:Disease Models, Animal; D007700:Kinetics; D018625:Microscopy, Atomic Force; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D055503:Protein Multimerization; D013696:Temperature; D014020:Tissue Extracts / Q000378:metabolism
D016229:Amyloid beta-Peptides; D029742:Caenorhabditis elegans Proteins; D010446:Peptide Fragments; D014020:Tissue Extracts; C078592:amyloid beta-protein (1-40); D010447:Peptide Hydrolases
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