Structure
Determination

The structure of N2C/D282C rhodopsin was solved by molecular replacement with the 1GZM coordinates of native rhodopsin [2]. The retinal and the sugar chains at position 2 and 15 had been removed from the search model. Yet already early maps obtained after rigid body refinement and density modification showed clear density for the ligand retinal, the sugar at position 15 and the engineered disulfide. The lack of the oligosaccharide normally liked to the mutated N2 further verified that we had found the right solution. Continued refinement and model building resulted in a clear map for most of the protein and a R-cryst and R-free of 0.287 and 0.325, respectively.

Retinal Omit map
Retinal Omit map (0.75 sigma) obtained after molecular replacement, rigid body refinement and density modification.

Top and side view of P31 crystal packing.
Top and side view of P31
crystal packing.

Refined 2Fo-Fc electron density map (0.9 sigma) of the N-terminal region.
Refined 2Fo-Fc electron density map (0.9 sigma) of the N-terminal region.


Comparison with Native Rhodopsin >


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