Three chaperones coordinate the breakup of amyloid fibrils in yeast riken_en nchembio
, Sedimentation coefficient distribution of Ssa1, Sis1, Sis1-HPD/AAA, Ssa1+Sis1 and Ssa1+Sis1-HPD/AAA at a concentration of 4 μM including 5 mM of ATP was obtained by AUC in 20C. Sedimentation was conducted at velocity of 42,000 rpm. Number of scans was 300 . overall distribution at a range of 0-42, Kinetics of accumulation of Ssa1-Alexa488 fluorescence on Sc4 amyloid in the presence of Sis1-Cy3 or Sis1 HPD/AAA-Cy3 with 5 mM ATP. 8 fibrils were analyzed from 2 observations for each condition.
5 μM Sc4 amyloid with 1 μM Hsp104 or Hsp104 mutants , 2 μM Ssa1 and 2 μM Sis1 was monitored at 30 °C by thioflavin T fluorescence. Data were normalized by the initial fluorescence intensities and averaged from 6 independent experiments . Data are presented as mean values +/− S.D., Representative TIRF images showing disaggregation of Sc4 amyloid-STELLA650 at various time points.
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