The 5 angstrom structure of heterologously expressed plant aquaporin SoPIP2;1
Kukulski, W ; Schenk, A D ; Johanson, Urban LU
; Braun, T
; de Groot, B L
; Fotiadis, D
; Kjellbom, Per
LU
and Engel, A
(2005)
In Journal of Molecular Biology
350(4).
p.611-616
- Abstract
- SoPIP2;1 is one of the major integral proteins in spinach leaf plasma membranes. In the Xenopus oocyte expression system its water channel activity is regulated by phosphorylation at the C terminus and in the first cytosolic loop. To assess its structure, SoPIP2;1 was heterologously expressed in Pichia pastoris as a His-tagged protein and in the non-tagged form. Both forms were reconstituted into 2D crystals in the presence of lipids. Tubular crystals and double-layered crystalline sheets of non-tagged SoPIP2;1 were observed and analyzed by cryo-electron microscopy. Crystalline sheets were highly ordered and diffracted electrons to a resolution of 2.96 angstrom. High-resolution projection maps of tilted specimens provided a 3D structure at... (More)
- SoPIP2;1 is one of the major integral proteins in spinach leaf plasma membranes. In the Xenopus oocyte expression system its water channel activity is regulated by phosphorylation at the C terminus and in the first cytosolic loop. To assess its structure, SoPIP2;1 was heterologously expressed in Pichia pastoris as a His-tagged protein and in the non-tagged form. Both forms were reconstituted into 2D crystals in the presence of lipids. Tubular crystals and double-layered crystalline sheets of non-tagged SoPIP2;1 were observed and analyzed by cryo-electron microscopy. Crystalline sheets were highly ordered and diffracted electrons to a resolution of 2.96 angstrom. High-resolution projection maps of tilted specimens provided a 3D structure at 5 angstrom resolution. Superposition of the SoPIP2;1 potential map with the atomic model of AQP1 demonstrates the generally well conserved overall structure of water channels. Differences concerning the extracellular loop A explain the particular crystal contacts between oppositely oriented membrane sheets of SoPIP2;1 2D crystals, and may have a function in rapid volume changes observed in stomatal guard cells or mesophyll protoplasts. This crystal packing arrangement provides access to the phosphorylated C terminus as well as the loop B phosphorylation site for studies of channel gating. (c) 2005 Elsevier Ltd. All rights reserved. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/152486
- author
- Kukulski, W
; Schenk, A D
; Johanson, Urban
LU
; Braun, T
; de Groot, B L
; Fotiadis, D
; Kjellbom, Per
LU
and Engel, A
- organization
- publishing date
- 2005
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Molecular Biology
- volume
- 350
- issue
- 4
- pages
- 611 - 616
- publisher
- Elsevier
- external identifiers
-
- wos:000230446000001
- pmid:15964017
- scopus:20544441912
- pmid:15964017
- ISSN
- 1089-8638
- DOI
- 10.1016/j.jmb.2005.05.001
- language
- English
- LU publication?
- yes
- id
- aeb2780b-8f7b-4f2a-a984-b54afec2acbc (old id 152486)
- date added to LUP
- 2016-04-01 15:43:36
- date last changed
- 2022-01-28 06:44:04
@article{aeb2780b-8f7b-4f2a-a984-b54afec2acbc,
abstract = {{SoPIP2;1 is one of the major integral proteins in spinach leaf plasma membranes. In the Xenopus oocyte expression system its water channel activity is regulated by phosphorylation at the C terminus and in the first cytosolic loop. To assess its structure, SoPIP2;1 was heterologously expressed in Pichia pastoris as a His-tagged protein and in the non-tagged form. Both forms were reconstituted into 2D crystals in the presence of lipids. Tubular crystals and double-layered crystalline sheets of non-tagged SoPIP2;1 were observed and analyzed by cryo-electron microscopy. Crystalline sheets were highly ordered and diffracted electrons to a resolution of 2.96 angstrom. High-resolution projection maps of tilted specimens provided a 3D structure at 5 angstrom resolution. Superposition of the SoPIP2;1 potential map with the atomic model of AQP1 demonstrates the generally well conserved overall structure of water channels. Differences concerning the extracellular loop A explain the particular crystal contacts between oppositely oriented membrane sheets of SoPIP2;1 2D crystals, and may have a function in rapid volume changes observed in stomatal guard cells or mesophyll protoplasts. This crystal packing arrangement provides access to the phosphorylated C terminus as well as the loop B phosphorylation site for studies of channel gating. (c) 2005 Elsevier Ltd. All rights reserved.}},
author = {{Kukulski, W and Schenk, A D and Johanson, Urban and Braun, T and de Groot, B L and Fotiadis, D and Kjellbom, Per and Engel, A}},
issn = {{1089-8638}},
language = {{eng}},
number = {{4}},
pages = {{611--616}},
publisher = {{Elsevier}},
series = {{Journal of Molecular Biology}},
title = {{The 5 angstrom structure of heterologously expressed plant aquaporin SoPIP2;1}},
url = {{http://dx.doi.org/10.1016/j.jmb.2005.05.001}},
doi = {{10.1016/j.jmb.2005.05.001}},
volume = {{350}},
year = {{2005}},
}