Extended Data Fig. 5: Photo-crosslinking experiments between NPY and Y1R.
From: Structural basis of ligand binding modes at the neuropeptide Y Y1 receptor
a, Mass spectra of photo-crosslinked Y1R with [Bpa1, K4[(Ahx)2-biotin]]NPY. Exemplary MALDI–TOF mass spectra of photo-crosslinked samples enzymatically digested by rLys-C and Glu-C. Potential Y1R fragments are labelled. Two independent experiments were performed with similar results. N, N terminus of Y1R (blue); E, ECL2 (red). b, Respective regions of NPY N terminus at Y1R. Amino acid sequence of Y1R with a C-terminal His-tag. The two detected regions within Y1R (N terminus (blue), ECL2 (red)) after crosslinking with [Bpa1,K4[(Ahx)2-biotin]]NPY are emphasized in boxes. The different sizes of the boxes represent different detected fragments (Extended Data Table 5). Experiments were repeated twice independently with similar results, and only fragments that were observed in both experiments are listed here and in Extended Data Table 5. c, Binding of Atto 520-labelled NPY (50 nM) to increasing amounts of cell-free produced Y1R in Brij-58. Data reflect fluorescence enhancement upon binding. An EC50 value of 69 nM was determined. Data shown are mean ± s.e.m. from six independent experiments performed in technical triplicate. c(Y1R), concentration of Y1R.
