Fig. 6: Concentration-dependent stoichiometry shift of ULK1C.
From: Structure and activation of the human autophagy-initiating ULK1C:PI3KC3-C1 supercomplex
Mass photometry measurements of (i) FIP200NTD, (ii) ULK1MIT:ATG13 (363–517), (iii) full-length PI3KC1-C1, (iv) FIP200NTD with truncated ULK1MIT:ATG13 (450–517; note: the molecular weight shifted because of the truncation), (v,vi) FIP200NTD with ULK1MIT:ATG13 (363–517) crosslinked at 10 nM in the absence or presence of 5 nM PI3KC3-C1, (vii,viii) FIP200NTD with ULK1MIT:ATG13 (363–517) crosslinked at 200 nM in the absence or presence of 100 nM PI3KC3-C1 and (ix,x) FIP200NTD with ULK1MIT:ATG13 (363–517) at crosslinked 1,000 nM in the absence or presence of 500 nM PI3KC3-C1. Solid curves represent fits to multiple Gaussian to estimate the average mass and the percentage of each population (Extended Data Table 1). The percentage was calculated from the area of the peaks in the range of 0–500 kDa. The schematic drawing illustrates the setting of each measurement (left) and the predicted assemblies (bottom). Left, the theoretical molecular weights of each assembly are shown next to the diagrams (i–iv). Right, the crosslinking concentrations of FIP200 and ULK1:ATG13 used for each measurement are indicated with red labels (v–x).
