Hydrogen bond analysis during MD simulation and mass spectrometry analysis of Unc5 peptides, related to Figure 2
(A–C) Quantification of MD simulation results for each of the four pseudo-symmetrical copies in the complex, for each of the three rUnc5D-hGPC3 interfaces described in Figure 2.
(D and E) Views of the electron density maps calculated for the X-ray crystal structure of Unc5Aiso1 (Seiradake et al., 2014): the 2Fo-Fc is shown in blue (1 sigma level). The Fo-Fc map is shown in red/green (+/− 3 sigma level). Extra density is observed on the first two of the TSP tryptophans of the consensus W1xxW2xxW3 motif.
(F–I) LCMSMS of the tryptic Unc5 peptides confirming the C-mannosylation of tryptophan residues in the TSP1 domains of Unc5 proteins expressed in HEK cells.