Protonation of Glutamate 208 Induces the Release of Agmatine in an Outward-facing Conformation of an Arginine/Agmatine Antiporter

• Published in: The Journal of biological chemistry 2011-06, Vol.286 (22), p.19693-19701
• Main Authors: Zomot, Elia, Bahar, Ivet
• Format: Article
• Language: English
• Subjects: Agmatine - chemistry; Agmatine - metabolism; Amino Acid Transport; Amino Acid Transport Systems - chemistry; Amino Acid Transport Systems - genetics; Amino Acid Transport Systems - metabolism; Antiporters - chemistry; Antiporters - genetics; Antiporters - metabolism; Binding Sites; Biological Transport - physiology; Computational Biology; Computer Modeling; Crystallography, X-Ray; Escherichia coli - chemistry; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins - chemistry; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Glutamic Acid - chemistry; Glutamic Acid - genetics; Glutamic Acid - metabolism; Membrane Proteins; Models, Molecular; Molecular Dynamics; Protein Conformation; Protein Structure; Protein Structure, Secondary
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M110.202085

Virulent enteric pathogens have developed several systems that maintain intracellular pH to survive extreme acidic conditions. One such mechanism is the exchange of arginine (Arg+) from the extracellular region with its intracellular decarboxylated form, agmatine (Agm2+). The net result of this process is the export of a virtual proton from the cytoplasm per antiport cycle. Crystal structures of the arginine/agmatine antiporter from Escherichia coli, AdiC, have been recently resolved in both the apo and Arg+-bound outward-facing conformations, which permit us to assess for the first time the time-resolved mechanisms of interactions that enable the specific antiporter functionality of AdiC. Using data from ∼1 μs of molecular dynamics simulations, we show that the protonation of Glu-208 selectively causes the dissociation and release of Agm2+, but not Arg+, to the cell exterior. The impact of Glu-208 protonation is transmitted to the substrate binding pocket via the reorientation of Ile-205 carbonyl group at the irregular portion of transmembrane (TM) helix 6. This effect, which takes place only in the subunits where Agm2+ is released, invites attention to the functional role of the unwound portion of TM helices (TM6 Trp-202–Glu-208 in AdiC) in facilitating substrate translocation, reminiscent of the behavior observed in structurally similar Na+-coupled transporters.