Folding, misfolding, and aggregation of polyglutamine peptides and proteins

BIOT 482

Matthew Tobelmann, tobelmann@wisc.edu, Christine Lee, Robert Walters, and Regina M. Murphy, regina@engr.wisc.edu. Department of Chemical and Biological Engineering, University of Wisconsin, 1415 Engineering Drive, Madison, WI 53706
In Huntington's and related diseases, proteins with expanded polyglutamine domains are expressed; these proteins aggregate into inclusions that are believed to cause neuronal degeneration. Synthetic polyglutamine peptides are useful models of polyglutamine-containing proteins. A widely accepted hypothesis is that polyglutamine aggregation follows a nucleation-elongation mechanism characterized by a lag time and a monomeric nucleus. We re-examined this hypothesis by measuring the aggregation kinetics of K2Q23K2, using light scattering and size exclusion chromatography. During the putative lag time there is substantial organization of the peptide into soluble linear aggregates. We propose that polyglutamine assembles first via hydrophobic interactions, which then convert into insoluble ß-sheet fibrils via slow conformational changes. Once fibrils form, monomer loss is accelerated, possibly through templated assembly. We established a model system using apomyoglobin as the host protein, and generated a library of mutants containing 16 - 102 glutamines. Detailed characterization of folding, stability, and aggregation of this library is underway.