Ammonia transporter

Ammonia transporters (TC# 1.A.11) are structurally related

The structure of the ammonia channel from E. coli,[3]^[4] was, at the time of its publication, the highest resolution structure of any integral membrane protein. It shows a trimer of subunits, each made up of 11 transmembrane segments (TMSs) and containing a pseudo two-fold symmetry.^[9] Each monomer contains a hydrophobic ammonia conducting channel.

While prokaryotic ammonia channel proteins have an N-terminal region which acts as a signal sequence and is cleaved in the mature protein,^[10] the Rhesus glycoproteins retain this as a 12th transmembrane helix in the mature protein.^[1]

Substrate specificity

Most functionally characterized members of the family are ammonium uptake transporters.^[11] Some, but not other Amt proteins also transport methylammonium.^[12][13] Detailed phylogenetic analyses of plant homologues have been published.^[14] In E. coli, NH₄⁺, rather than NH₃, may be the substrate of AmtB, but controversy still exists.^[15][16][17] If NH₄⁺ is transported, K⁺ possibly serves as a counter ion in an antiport process with K⁺, and that one histidine removes a proton off of NH₄⁺ to yield NH₃.^[15]

Transport reaction

The generalized transport reaction catalyzed by members of the Amt family are suggested to be:

NH₄⁺ (out) ⇌ NH₄⁺ (in)

Mechanism

The X-ray structures have revealed that the pore of the Amt and Rh proteins is characterized by a hydrophobic portion about 12 Å long, in which electronic density was observed in the crystallographic study of AmtB from Escherichia coli. This electronic density was initially only observed when crystals were grown in the presence of ammonium, and was thus attributed to ammonia molecules. The Amt/Rh protein mechanism might involve the single-file diffusion of NH₃ molecules. However, the pore could also be filled with water molecules. The possible presence of water molecules in the pore lumen calls for a reassessment of the notion that Amt/Rh proteins work as plain NH₃ channels. Indeed, functional experiments on plant ammonium transporters and Rh proteins suggest a variety of permeation mechanisms including the passive diffusion of NH₃, the antiport of NH₄⁺/H⁺, the transport of NH₄⁺, or the cotransport of NH₃/H⁺. Lamoureux et al. discuss these mechanisms in light of functional and simulation studies on the AmtB transporter.^[18]

Regulation

In E. coli the AmtB gene is expressed only under limiting nitrogen levels to yield the AmtB protein. It is co-expressed with the GlnK gene which encodes a PII protein. This protein is also trimeric and remains in the cytoplasm.^[19] It is covalently modified by a U/U deuridylylated group at Y51. The hydrolyzed product, adenosine 5'-diphosphate, orients the surface of GlnK for AmtB blockade. When nitrogen levels outside the cell rise, the ammonia channel must be deactivated to prevent excessive ammonia entering the cell (where ammonia would be combined with glutamate to make glutamine, utilizing ATP and thereby depleting the cell's ATP reserves). This deactivation is achieved by deuridylylation of the GlnK protein which then binds to the cytoplasmic face of AmtB and inserts a loop into the ammonia conducting pore. At the tip of this loop is an arginine residue which sterically blocks the channel.^[20]

Human ammonia transporter-related proteins

RHAG, RHBG, RHCE, RHCG, RHD

References