Extended Data Fig. 6: Cocaine binding to purified hDATTwin is similar to hDAT expressed in COS7 cells. | Nature

Extended Data Fig. 6: Cocaine binding to purified hDATTwin is similar to hDAT expressed in COS7 cells.

From: Structure of the human dopamine transporter in complex with cocaine

Extended Data Fig. 6

a, Inhibition of [3H]MFZ 2-12 binding by cocaine, determined in both intact COS7 cells expressing hDATWT (blue) and in detergent solubilized hDATTwin (purple). The affinity (Ki) determined for cocaine is similar in both systems (Extended Data Fig. 1d). Data are shown as individual data points, n = 3 (detergent solubilized hDATTwin) and n = 4 (hDATTwin expressed in COS7 cells) biological replicates, performed in three technical replicates. Curves are fitted to the average of the data points. b, To the left, the obtained cryo-EM density of the hDATTwin:cocaine complex contoured at 12σ and colored according to local resolution. In the middle, a cut through of the density exposing a non-proteinaceous density, marked by a dashed box, to which R-cocaine was fitted (shown to the right). c, Diagram showing the type of interactions between the functional groups of cocaine and involved hDATTwin residues (diagram calculated and generated by Maestro, Schrodinger suite). Residues within 6 Å distance are shown as teardrops, with the pointy edge representing side chain directionality. The backbone is represented as a black line connecting residues. Residues of the sequence not within the 6 Å distance are shown as black dots. Residues that do not have a line drawn for an interaction are involved in nonspecific hydrophobic interactions with cocaine. The binding pocket is represented by a line around the ligand, colored according to the nearest protein residue’s property. Solvent exposed areas are indicated on the ligand atoms, and by the break in the line drawn around the pocket.

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