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Stoichiometry and Substrate Affinity of the Mannitol Transporter, EnzymeII^sup mtl^, from Escherichia coli
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Biophysical Journal
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July 1, 2005
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Copyright informationCopyright Biophysical Society Jul 2005. Provided by ProQuest LLC. (Hide copyright information)
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ABSTRACT
Uptake and consecutive phosphorylation of mannitol in Escherichia coli is catalyzed by the mannitol permease Enzymell^sup mtl^. The substrate is bound at an extracellular-oriented binding site, translocated to an inward-facing site, from where it is phosphorylated, and subsequently released into the cell. Previous studies have shown the presence of both a high- and a lowaffinity binding site with K^sub D^-values in the nano- and micromolar range, respectively. However, reported K^sub D^-values in literature are highly variable, which casts doubts about the reliability of the measurements and data analysis. Using an optimized binding measurement system, we investigated the ...
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