Fig. 5: Confirmation of the TCR–CD3 resting conformation in vivo.
From: The resting and ligand-bound states of the membrane-embedded human T-cell receptor–CD3 complex
a Comparison of Cα-Cα distances between apposing faces of CD3δ and TCRα in GDN and ND-I, highlighting plausible engineered disulfide positions in ND-I (distances in blue shades) that are not geometrically plausible in GDN (distances in warm shades). b Non-reducing SDS-PAGE of whole-cell lysates from HEK cells infected with the indicated TCR–CD3 virus genotypes Western blotted for EGFP-tagged TCRβ. Engineered disulfides yielding a TCRαβ–CD3δ covalent adduct resulted in an upward gel shift (asterisks). Color coding and distances as in (a). Full blots from two experimental replicates are shown in Supplementary Fig. 5b. Design of intrachain disulfide bonds in the closed conformation of TCRα (c) and TCRβ (d). Distances color-coded as above. Mass spectrometric detection of intrachain disulfide bonds in TCRα (e) and TCRβ (f). EThcD fragment spectra of disulfide-linked dipeptides shows gas-phase reduced linear peptides (red, peptide A; blue, peptide B; reduced Cys, bold) and their fragment ions (magenta, peptide A; cyan, peptide B). Charge-reduced precursor ions are in gray.
