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Fig. 2. Mapping of the binding sites of Pontin and Reptin in Hint1 and vice versa. (A) C-terminal and N-terminal deletion constructs of Pontin were expressed as GST-fusion proteins as schematically presented and used to map the binding site of Hint1 in Pontin to amino acids 214-294 (lanes 2,4,10,12,14) in pull-down assays. (B) Hint1 binds to amino acids 218-289 in Reptin (lanes 4,10,14,16,18) as mapped with C- and N-terminally deleted MBP-Reptin fusion proteins. (C) MBP-Pontin and -Reptin bind to the N-terminus of Hint1 in pull-down assays with GST-Hint1{Delta}C (amino acids 1-72) (lanes 5 and 8). The C-terminal half (amino acids 66-126) of Hint1 (GST-Hint1{Delta}N) mediates dimerization (lane 12).





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