Mechanism of ammonia transport by Amt/MEP/Rh: structure of AmtB at 1.35 A

Science. 2004 Sep 10;305(5690):1587-94. doi: 10.1126/science.1101952.

Abstract

The first structure of an ammonia channel from the Amt/MEP/Rh protein superfamily, determined to 1.35 angstrom resolution, shows it to be a channel that spans the membrane 11 times. Two structurally similar halves span the membrane with opposite polarity. Structures with and without ammonia or methyl ammonia show a vestibule that recruits NH4+/NH3, a binding site for NH4+, and a 20 angstrom-long hydrophobic channel that lowers the NH4+ pKa to below 6 and conducts NH3. Favorable interactions for NH3 are seen within the channel and use conserved histidines. Reconstitution of AmtB into vesicles shows that AmtB conducts uncharged NH3.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Ammonia / metabolism*
  • Binding Sites
  • Biological Transport
  • Cation Transport Proteins / chemistry*
  • Cation Transport Proteins / genetics
  • Cation Transport Proteins / metabolism
  • Cell Membrane / chemistry
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / chemistry*
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Hydrogen Bonding
  • Hydrogen-Ion Concentration
  • Hydrophobic and Hydrophilic Interactions
  • Liposomes
  • Membrane Potentials
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Protein Folding
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • Quaternary Ammonium Compounds / metabolism
  • Rh-Hr Blood-Group System / chemistry
  • Rh-Hr Blood-Group System / metabolism
  • Sequence Alignment
  • Water / chemistry
  • Water / metabolism

Substances

  • AmtB protein, E coli
  • Cation Transport Proteins
  • Escherichia coli Proteins
  • Liposomes
  • Quaternary Ammonium Compounds
  • Rh-Hr Blood-Group System
  • Water
  • Ammonia

Associated data

  • PDB/1U77
  • PDB/1U7C
  • PDB/1U7G