Figure EV4. Competitive uptake assay of L‐[³H]Leu by WT or mutated variants of MhsT in the presence or absence of selected amino acids.

• A–J
  MhsT‐WT uptake of L‐leucine is inhibited to various degree by all of the hydrophobic amino acids. The M236F mutant shows complete inability to compete l‐leucine with hydrophobic amino acids, indicating impaired binding or transport. The triple18 variant harbors the following substitutions: L328T, M236F, and V331A. Uptake measurements were performed in Escherichia coli strain YG228 expressing the indicated variants. Data are the mean ± SEM of two independent experiments each performed as technical duplicates.