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

Published

Journal Article

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 approximately 80 A 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 A resolution. Cryoelectron microscopy provided a 3.7 A 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.

Full Text

Duke Authors

Cited Authors

  • Braun, T; Philippsen, A; Wirtz, S; Borgnia, MJ; Agre, P; Kühlbrandt, W; Engel, A; Stahlberg, H

Published Date

  • August 2000

Published In

Volume / Issue

  • 1 / 2

Start / End Page

  • 183 - 189

PubMed ID

  • 11265760

Pubmed Central ID

  • 11265760

International Standard Serial Number (ISSN)

  • 1469-221X

Digital Object Identifier (DOI)

  • 10.1038/sj.embor.embor615

Language

  • eng

Conference Location

  • England