The interplay between PolyQ and protein context delays aggregation by forming a reservoir of protofibrils.
Polyglutamine (polyQ) diseases are inherited neurodegenerative disorders caused by the expansion of CAG codon repeats, which code for polyQ in the corresponding gene products. These diseases are associated with the presence of amyloid-like protein aggregates, induced by polyQ expansion. It has been suggested that the soluble aggregates rather than the mature fibrillar aggregates are the toxic species, and that the aggregation properties of polyQ can be strongly modulated by the surrounding protein context. To assess the importance of the protein carrier in polyQ aggregation, we have studied the misfolding pathway and the kinetics of aggregation of polyQ of lengths above (Q41) and below (Q22) the pathological threshold fused to the well-characterized protein carrier glutathione S-transferase (GST). This protein, chosen as a model system, is per se able to misfold and aggregate irreversibly, thus mimicking the behaviour of domains of naturally occurring polyQ proteins. We prove that, while it is generally accepted that the aggregation kinetics of polyQ depend on its length and are faster for longer polyQ tracts, the presence of GST alters the polyQ aggregation pathway and reverses this trend. Aggregation occurs through formation of a reservoir of soluble intermediates whose populations and kinetic stabilities increase with polyQ length. Our results provide a new model that explains the toxicity of expanded polyQ proteins, in which the interplay between polyQ regions and other aggregation-prone domains plays a key role in determining the aggregation pathway.
Bulone
Donatella
D
National Research Council (CNR), Istituto di Biofisica di Palermo, Palermo, Italy.
Masino
Laura
L
Thomas
David J
DJ
San Biagio
Pier Luigi
PL
Pastore
Annalisa
A
D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D002942:Circular Dichroism; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D020271:Heredodegenerative Disorders, Nervous System / Q000235:genetics / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008027:Light; D046529:Microscopy, Electron, Transmission; D008954:Models, Biological; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D055503:Protein Multimerization; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D012542:Scattering, Radiation; D012995:Solubility; D013816:Thermodynamics; D019680:Trinucleotide Repeat Expansion
D002352:Carrier Proteins; D046912:Multiprotein Complexes; D010455:Peptides; D011993:Recombinant Fusion Proteins; C097188:polyglutamine; D005982:Glutathione Transferase