Journal: bioRxiv
Article Title: Local aromatic interactions define temperature sensitivity of phase separation in an intrinsically disordered protein
doi: 10.64898/2026.05.07.723405
Figure Lengend Snippet: ( A ) Schematic representation of Tyr-substituted Sup35NM mutants (YA-NM, YAh-NM, YF-NM, and YFh-NM). Tyr positions are indicated by black lines, Ala substitutions by red lines, and Phe substitutions by green lines. ( B ) Temperature-dependent turbidity profiles of Tyr-to-Ala Sup35NM mutants (YA-NM and YAh-NM) in 20% (w/v) PEG 20,000. ( C ) Schematic representation of Tyr-substituted Sup35N mutants (YA-N, YAh-N, YF-N, and YFh-N). ( D ) Temperature-dependent turbidity profiles of Tyr-to-Ala Sup35N mutants, YA-N and YAh-N in 20% (w/v) PEG 20,000. ( E ) Blue-Native PAGE analysis of Sup35NM mutants carrying Tyr-to-Ala (red) or Tyr-to-Phe (green) substitutions. ( F ) Temperature-dependent turbidity profiles of Tyr-to-Phe Sup35NM mutants (YF-NM and YFh-NM) in 20% (w/v) PEG 20,000. ( G ) Temperature-dependent turbidity profiles of Tyr-to-Phe Sup35N mutants (YF-N and YFh-N) in 20% (w/v) PEG 20,000. ( H ) (left) Representative optical microscopy images showing droplet morphology of YF-NM and YF-N mutants at 20°C in 20% (w/v) PEG 20,000. Scale bars, 10 µm. Right panels show magnified views. Scale bars, 5 μm.
Article Snippet: The final reaction mixture was adjusted to contain 10 μM protein, 10 mM sodium phosphate buffer (pH 7.0), and 10–20% (w/v) polyethylene glycol (PEG, average molecular weight 20,000, Nacalai Tesque).
Techniques: Blue Native PAGE, Microscopy