Skip to main content
. 2012 Oct 17;103(8):1706–1718. doi: 10.1016/j.bpj.2012.09.001

Figure 3.

Figure 3

Biophysical properties of recombinant inter-CBS linker protein. (A) Prediction of disorder in the inter-CBS linker domain using PONDR (25). Predictions were generated for WT and S742E/S747E linkers. PONDR scores were identical for the two proteins except for amino acids 728–760, which are shown on the plot and contain the two regulatory phosphorylation sites (S742 and S747 shown in red). In the WT linker (black line), amino acids 734–753 have near-zero PONDR scores suggesting a high degree of order. Replacement of S742 and S747 with glutamate (red line) to mimic phosphorylation increases the PONDR scores in this region suggesting an increase in disorder. The GCK-3-binding motif (RFLI; amino acids 668–671) is shown. (B) SDS-PAGE gel of 1 μg of purified inter-CBS linker protein (amino acids 619–793). (C) Near ultraviolet CD spectra of WT and S742E/S747E mutant linkers. Spectra indicate that the WT and S742E/S747E mutant linkers contain little secondary structure.