|
|
|
|
|
Aquaporin Structure Elucidates Water Transport
From aqueducts to osmosis, water transport is crucial to life. Yet, precisely
how life manages the transport of water across membranes has remained
a mystery for eonsuntil now. A team of researchers from the Berkeley
Lab Life Sciences Division has solved the structure of aquaporin-1 (AQP1),
a membrane protein that controls the movement of water molecules into
and out of mammalian cells. It is a member of the aquaporin superfamily,
whose members transport water or water and glycerol or urea. The new structure
offers a resolution of 2.2 Å, allowing researchers to deduce just how
the protein does its job.
After preliminary work at the National Synchrotron Light Source
and the Stanford Synchrotron Radiation Laboratory, the team turned
to the Berkeley Center for Structural Biology and ALS Beamline
5.0.2. They began by studying thallium-derivatized crystals of
AQP1 from bovine red blood cells by multiwavelength anomalous
diffraction (MAD). They then refined the resulting data set to
2.2 Å by using crystals grown in the presence of gold cyanide.
The high-resolution structure shows atomic-level details of the
protein and water molecules captured in transit.
The AQP1 tetramer viewed looking down the pores from the
cytoplasmic side, normal to the membrane. One monomer of
the four is represented as a solid space-filling model. |
The overall structure of AQP1 is that of a tetramer, the four parts
(monomers) of which each define a single pore. These monomers are
arranged side by side in a tight cluster, with the pores running
parallel. Each monomer in turn comprises six membrane-spanning helices
that partially surround two shorter helices. The short non-membrane-spanning
helices make up the major portion of the pore. Each pore has a dumbbell-like
shape. One broad end is the cytoplasmic vestibule; the other is
the extracellular vestibule. The bar of the dumbbell is the selectivity
filter, which narrows to a constriction region on the extracellular
end.
|
More Than Just a Bag of Water
|
In the new structure, the key elements of the constriction region
can be discerned. A series of carbonyl oxygens forms a hydrophilic
path across the region and through the rest of the selectivity filter.
One of these oxygens, along with a histidine residue and an arginine
residue, forms the hydrophilic face of the constriction region.
Opposite this face is a hydrophobic face formed by a phenylalanine
residue. Three of the four residues that form the constriction region
(the arginine, histidine, and phenylalanine residues) are conserved
in all known water-specific aquaporins. This observation suggests
that the presence of these residues can be used as a marker for
identifying other water-specific aquaporins. |
|
|
Side view of AQP1 showing the pore profile (turquoise
dots) and the residues that line the pore (opaque ball-and-stick
structures). The extracellular vestibule is above; cytoplasmic,
below. The pinched-in area with the highest concentration of
turquoise dots is the constriction region. |
The hydrophilic path across the selectivity filter,
highlighted by ball-and-stick structures of the side chains
involved. The green spheres are the water molecules observed
in transit. The constriction region is indicated by the blue
arrow. |
An
earlier study by a different groupalso done at the ALSrevealed
the structure of a closely related bacterial channel, the Escherichia
coli glycerol facilitator (GlpF), which selectively transports glycerol.
The structure found for GlpF differs from that of AQP1 in that its
constriction region is about 1 Å wider and slightly less polar.
The resulting decreases in steric hindrance (physical blocking)
and hydrophilicity favor glycerol transport at the expense of rapid
water throughput. Despite the functional difference, the GlpF constriction
region is strikingly similar to its AQP1 counterpart. It differs
only in the replacement of a histidine by a glycine and a cysteine
by a phenylalanine. As both the cysteine and the phenylalanine provide
carbonyl oxygens to shape the constriction region, the difference
in functionality turns out to depend upon the residue found in the
location of the histidine (H182) in AQP1. This residue thus appears
to be key in defining selectivity throughout the aquaporin superfamily.
Research conducted by H. Sui, B.-G. Han, J.K. Lee, P. Walian, and
B.K. Jap (Berkeley Lab).
Research funding: National Institutes of Health; U.S. Department
of Energy, Office of Health and Environmental Research. Operation
of the ALS is supported by the U.S. Department of Energy, Office
of Basic Energy Sciences.
Publication about this research: H. Sui, B.-G. Han, J.K. Lee, P.
Walian, and B.K. Jap, "Structural basis of water-specific transport
through the AQP1 water channel," Nature 414, 878 (2001).
ALSNews
Vol. 210, October 30, 2002
More ALS Science
|
|
|