Figure 8
Overlay of sections of the [1H-15N] HSQC spectra (Supplementary Fig. S11) of [U-13C,15N] LC3C before (black) and 3.5 h (red) and 7.0 h (green) after incubation with PKA to monitor the progress of in vitro phosphorylation. (A) Intense resonance signals of Met1 and Lys6 did not shift, while the shift of the resonance of Glu25 could be traced (open arrow) and the resonance of Arg22 disappears. (B) The populations of the split signal of Phe13 in the unphosphorylated state (black) shifted towards the minor peak position (filled arrow), while the resonance of Val26 could be traced to a new peak position (open arrow). (C) The intensities of the resonances of Ser18 and Ala20 decreased within 3.5 h and could not be traced to newly emerging peaks in the vicinity, in contrast to Glu123 (filled arrow) and (D) Glu111. The resonance of Met97 was not influenced by PKA reaction progress. (E) Split backbone amide resonances (magenta), traceable backbone amide resonances (cyan), and disappearing backbone amide resonances (orange) after incubation with PKA color-coded onto a schematic representation of the secondary structure of the lowest-energy solution structure of LC3C (same view as in Fig. 5). The figure was drawn with MolScript 2.1.2100 and rendered with Raster3D 3.0101.
