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

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 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.

journal_name

EMBO Rep

journal_title

EMBO reports

authors

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

doi

10.1093/embo-reports/kvd022

keywords:

subject

Has Abstract

pub_date

2000-08-01 00:00:00

pages

183-9

issue

2

eissn

1469-221X

issn

1469-3178

pii

embor615

journal_volume

1

pub_type

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