Oxidative cross-linking stabilizes C-D2R-D2RV hetero-oligomers via Cys168.
A, immunoblot probed with anti-GFP primary antibody of lysates from cells
transfected with equal amounts of C-D2R or D2R-V (both Cys168 or Ser168)
plasmid DNA, with or without CuP treatment; the predicted molecular mass of
both protomers is ∼80 kDa. B and C, recovery of D2R-V fluorescence after
antibody cross-linking alone (anti-GFP) or antibody and oxidative
cross-linking (+CuP); lines represent the mean ± S.E.M. of all cells
expressing C-D2R Cys168 and D2R-V Cys168 (B) or C-D2R Cys168 and D2R-V Ser168
(C). D to F, recovery of normalized fluorescence for cells expressing
(measured at time = 180 s) for C-D2R Cys168 and D2R-V Cys168 (D; control,
n = 46; CuP, n = 48), C-D2R Cys168 and D2R-V Ser168 (E;
control, n = 25; CuP, n = 31), and C-D2R Ser168 and D2R-V
Cys168 (F; control, n = 11; CuP, n = 17); **,
P < 0.001; n.s., P > 0.1; bars represent mean ±
S.E.M.