Figure 1.

Reactivation of wild-type SERT and cysteine replacement mutants. (A) Cysteine-dependent reactivation. Confluent INT-407 cells in 24-well plates were infected with T7 vaccinia virus and transfected with cDNA encoding wild-type SERT and mutants I172C, I179C, and L564C. After incubation for 16–20 h at 37°C cells were treated with MTSET for 10 min, rinsed with PBS, treated with 12 mM cysteine for the indicated time, and assayed for transport activity. (B) Spontaneous reactivation. Cells expressing I172C, I179C, and L564C were inactivated with MTSET as described above and incubated in the absence of cysteine for the indicated time before assaying transport.