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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^NTQ₃₀P₆ (F17W) by multiple analyses. (A) Fluorescence emission maximum of Trp residue at position 17 in resuspended aggregates isolated from reaction of htt^NTQ₃₀P₆ (F17W) (■) or htt^NTQ₃ (F17W) (Δ). The emission maximum of monomeric peptide (•) is plotted as being equivalent to that of initial aggregates, since this is the result obtained for the F17W mutant of the shorter, less rapidly aggregating htt^NTQ₂₀P₁₀. The htt^NT aggregation reaction was carried out to 800 hrs, at which time W17 remained completely solvent exposed (not shown). (B) Time course monitored by HPLC sedimentation assay (——◆——), R²=0.983, S.E.= ±6.0), thioflavin T fluorescence (——Δ——, R²=0.994, S.D.= ±3.9) and right angle light scattering (---○---, R²=0.983, S.D.= ±6.0). Inset, first 10 hrs. (C) Dot blots of htt^NTQ₃₀P₆ (F17W) time points using the antibody MW1. Top row: unfractionated aliquots of the reaction mixture (time in hrs; M = non-incubated monomer). Bottom row: equivalent masses of isolated aggregates (no material in the “M” column in this row).

Time course of aggregation of htt^NTQ₃₀P₆ (F17W) by multiple analyses. (A) Fluorescence emission maximum of Trp residue at position 17 in resuspended aggregates isolated from reaction of htt^NTQ₃₀P₆ (F17W) (■) or htt^NTQ₃ (F17W) (Δ). The emission maximum of monomeric peptide (•) is plotted as being equivalent to that of initial aggregates, since this is the result obtained for the F17W mutant of the shorter, less rapidly aggregating htt^NTQ₂₀P₁₀. The htt^NT aggregation reaction was carried out to 800 hrs, at which time W17 remained completely solvent exposed (not shown). (B) Time course monitored by HPLC sedimentation assay (——◆——), R²=0.983, S.E.= ±6.0), thioflavin T fluorescence (——Δ——, R²=0.994, S.D.= ±3.9) and right angle light scattering (---○---, R²=0.983, S.D.= ±6.0). Inset, first 10 hrs. (C) Dot blots of htt^NTQ₃₀P₆ (F17W) time points using the antibody MW1. Top row: unfractionated aliquots of the reaction mixture (time in hrs; M = non-incubated monomer). Bottom row: equivalent masses of isolated aggregates (no material in the “M” column in this row).