The 4.5 A structure of human AQP2.
Located in the principal cells of the collecting duct, aquaporin-2 (AQP2) is responsible for the regulated water reabsorption in the kidney and is indispensable for the maintenance of body water balance. Disregulation or malfunctioning of AQP2 can lead to severe diseases such as nephrogenic diabetes insipidus, congestive heart failure, liver cirrhosis and pre-eclampsia. Here we present the crystallization of recombinantly expressed human AQP2 into two-dimensional protein-lipid arrays and their structural characterization by atomic force microscopy and electron crystallography. These crystals are double-layered sheets that have a diameter of up to 30 microm, diffract to 3 A(-1) and are stacked by contacts between their cytosolic surfaces. The structure determined to 4.5 A resolution in the plane of the membrane reveals the typical aquaporin fold but also a particular structure between the stacked layers that is likely to be related to the cytosolic N and C termini.
Schenk
Andreas D
AD
M. E. Müller Institute for Microscopy, Biozentrum, University of Basel, Klingelbergstrasse 70, 4056 Basel, Switzerland. andreas.engel@unibas.ch
Werten
Paul J L
PJ
Scheuring
Simon
S
de Groot
Bert L
BL
Müller
Shirley A
SA
Stahlberg
Henning
H
Philippsen
Ansgar
A
Engel
Andreas
A
D051399:Aquaporin 2; D020346:Aquaporins / Q000737:chemistry* / Q000648:ultrastructure*; D020285:Cryoelectron Microscopy; D003460:Crystallization; D006801:Humans; D018625:Microscopy, Atomic Force; D017348:Microscopy, Electron, Scanning Transmission; D046529:Microscopy, Electron, Transmission
C496023:AQP2 protein, human; D051399:Aquaporin 2; D020346:Aquaporins