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. Author manuscript; available in PMC: 2009 Oct 1.

Published in final edited form as: Nat Struct Mol Biol. 2009 Mar 8;16(4):380–389. doi: 10.1038/nsmb.1570

Time course of aggregation of htt^NTQ₂₀P₁₀ by multiple analyses. (A) Trypsin sensitivity of either monomer (t = 0) or aggregates isolated by centrifugation at either 42 or 700 hours (see Methods). (B) Properties of isolated aggregates: fluorescence emission maxima of Trp residues in the mutant peptides F11W (——•——, R²=0.994, S.D.= ±0.5) and F17W (····▲····, R²=0.916, S.D.= ±1.2); elongation rate constants for biotinyl-Q₂₉ for isolated aggregates adherent to microtiter plate wells (---□---, R²=0.748, S.D.= ±0.19). (C) Overall aggregation kinetics of WT peptide monitored by the HPLC sedimentation assay (---◆---, R²=0.992, S.D.= ±3.1) and by ThT fluorescence (——Δ——, R²=0.974, S.D.= ±6.0). (D) Dot blot of non-incubated monomer (M) and isolated aggregates developed with to the anti-polyQ MW1 antibody. (E) Fourier transform infrared (FTIR) spectra of aggregates. Monomeric Q₁₅ (a); aggregates of htt^NTQ₂₀P₁₀ (F17W) isolated at 45 hrs (b), 120 hrs (c), and 120 days (d); aggregates of htt^NTQ₃₆P₁₀ isolated at 7 days (e); aggregates of Q₃₀ isolated at 30 days (f). Amide I frequency values normally assigned to secondary structural features ⁵⁹ and Gln side chains ⁶⁰ are shown with bars at the top of the panel.

Time course of aggregation of htt^NTQ₂₀P₁₀ by multiple analyses. (A) Trypsin sensitivity of either monomer (t = 0) or aggregates isolated by centrifugation at either 42 or 700 hours (see Methods). (B) Properties of isolated aggregates: fluorescence emission maxima of Trp residues in the mutant peptides F11W (——•——, R²=0.994, S.D.= ±0.5) and F17W (····▲····, R²=0.916, S.D.= ±1.2); elongation rate constants for biotinyl-Q₂₉ for isolated aggregates adherent to microtiter plate wells (---□---, R²=0.748, S.D.= ±0.19). (C) Overall aggregation kinetics of WT peptide monitored by the HPLC sedimentation assay (---◆---, R²=0.992, S.D.= ±3.1) and by ThT fluorescence (——Δ——, R²=0.974, S.D.= ±6.0). (D) Dot blot of non-incubated monomer (M) and isolated aggregates developed with to the anti-polyQ MW1 antibody. (E) Fourier transform infrared (FTIR) spectra of aggregates. Monomeric Q₁₅ (a); aggregates of htt^NTQ₂₀P₁₀ (F17W) isolated at 45 hrs (b), 120 hrs (c), and 120 days (d); aggregates of htt^NTQ₃₆P₁₀ isolated at 7 days (e); aggregates of Q₃₀ isolated at 30 days (f). Amide I frequency values normally assigned to secondary structural features ⁵⁹ and Gln side chains ⁶⁰ are shown with bars at the top of the panel.