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DA uptake initiated to determine the degree of transport inactivation by MTSET. Importantly, response of an inserted cysteine on this place (I159C) with MTSET ends in inactivation of the DAT permitting the use of DA uptake as a functional learn-out for I159C reactivity (36). The I159C mutant was generated in a MTSET insensitive DAT background (E2C) by which the two external endogenous cysteines had been mutated to alanines (C90A-C306A). The second assay relies on the reactivity of a cysteine inserted into position 159 in TM3 of the DAT positioned in the vicinity of the ligand binding site on the extracellular aspect. The predicted binding modes of R-modafinil and S-modafinil in the central binding site of DAT, respectively. Herein we compare cocaine to the clinically out there (±) modafinil, its R-enantiomer (armodafinil) and S-modafinil at the DAT. Studies with the Y335A DAT mutant showed that the R- and S-enantiomers tolerated the inward going through conformation better than cocaine, which was further supported by MTSET reactivity on the DAT E2C I159C.


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