<p>(A) Ribbon structure of Guk1 (PDB 1EX7). Positions of the four missense mutations and predicted ∆∆G values are indicated. Loss of fluorescence measured by flow cytometry after a two hour incubation at 37°C with cycloheximide is indicated in brackets. (B) Cellular thermal shift assay of Guk1 and Guk1-7 fused to a six histidine tag in lysates derived from cells grown at 25°C. One representative anti-His Western Blot is shown. The graph represents the means and standard deviations of Guk1 levels from three independent experiments. (C) Guk1 and Guk1-7 fused to a six histidine tag were expressed in cells grown at 25°C or shifted to 37°C for 20 min. Total cell lysate (T), soluble (S), and pellet fractions (P) were immunoblotted with anti-His antibodies.</p
