The 3.7 Å projection map of the glycerol facilitator GlpF: a variant of the aquaporin tetramer

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The 3.7 Å projection map of the glycerol facilitator GlpF: a variant of the aquaporin tetramer

Publisher
Oxford University Press
Publication Date
Aug 15, 2000
Source
PMC
Keywords
Disciplines
  • Biology
License
Unknown

Abstract

GlpF, the glycerol facilitator protein of Escherichia coli, is an archetypal member of the aquaporin superfamily. To assess its structure, recombinant histidine-tagged protein was overexpressed, solubilized in octylglucoside and purified to homogeneity. Negative stain electron microscopy of solubilized GlpF protein revealed a tetrameric structure of ∼80 Å side length. Scanning transmission electron microscopy yielded a mass of 170 kDa, corroborating the tetrameric nature of GlpF. Reconstitution of GlpF in the presence of lipids produced highly ordered two-dimensional crystals, which diffracted electrons to 3.6 Å resolution. Cryoelectron microscopy provided a 3.7 Å projection map exhibiting a unit cell comprised of two tetramers. In projection, GlpF is similar to AQP1, the erythrocyte water channel. However, the major density minimum within each monomer is distinctly larger in GlpF than in AQP1.

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