The Molecular Basis of K+ Exclusion by the Escherichia coli Ammonium Channel AmtB

Members of the Amt family of channels mediate the transport of ammonium. The form of ammonium, NH3 or NH4+, carried by these proteins remains controversial, and the mechanism by which they select against K+ ions is unclear. We describe here a set of Escherichia coli AmtB proteins carrying mutations at the conserved twin-histidine site within the conduction pore that have altered substrate specificity and now transport K+. Subsequent work established that AmtB-mediated K+ uptake occurred against a concentration gradient and was membrane potential-dependent. These findings indicate that the twin-histidine element serves as a filter to prevent K+ conduction and strongly support the notion that Amt proteins transport cations (NH4+ or, in mutant proteins, K+) rather than NH3 gas molecules through their conduction pores.