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Affinity tags can reduce merohedral twinning of membrane protein crystals

Backmark, Anna; Nyblom, Maria; Tornroth-Horsefield, Susanna; Kosinska-Eriksson, Urszula; Nordén, Kristina LU ; Agemark, Maria LU ; Kjellbom, Per LU ; Johanson, Urban LU ; Hedfalk, Kristina and Lindkvist- ersson, Karin LU , et al. (2008) In Acta Crystallographica. Section D: Biological Crystallography 64. p.1183-1186
Abstract
This work presents a comparison of the crystal packing of three eukaryotic membrane proteins: human aquaporin 1, human aquaporin 5 and a spinach plasma membrane aquaporin. All were purified from expression constructs both with and without affinity tags. With the exception of tagged aquaporin 1, all constructs yielded crystals. Two significant effects of the affinity tags were observed: crystals containing a tag typically diffracted to lower resolution than those from constructs encoding the protein sequence alone and constructs without a tag frequently produced crystals that suffered from merohedral twinning. Twinning is a challenging crystallographic problem that can seriously hinder solution of the structure. Thus, for integral membrane... (More)
This work presents a comparison of the crystal packing of three eukaryotic membrane proteins: human aquaporin 1, human aquaporin 5 and a spinach plasma membrane aquaporin. All were purified from expression constructs both with and without affinity tags. With the exception of tagged aquaporin 1, all constructs yielded crystals. Two significant effects of the affinity tags were observed: crystals containing a tag typically diffracted to lower resolution than those from constructs encoding the protein sequence alone and constructs without a tag frequently produced crystals that suffered from merohedral twinning. Twinning is a challenging crystallographic problem that can seriously hinder solution of the structure. Thus, for integral membrane proteins, the addition of an affinity tag may help to disrupt the approximate symmetry of the protein and thereby reduce or avoid merohedral twinning. (Less)
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publication status
published
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Acta Crystallographica. Section D: Biological Crystallography
volume
64
pages
1183 - 1186
publisher
International Union of Crystallography
external identifiers
  • wos:000260197900012
  • scopus:56649092937
ISSN
1399-0047
DOI
10.1107/S090744490802948X
language
English
LU publication?
yes
id
c67e4246-3ec7-4d1a-aac0-c11a2f1f0116 (old id 1284820)
date added to LUP
2009-02-09 09:00:02
date last changed
2017-02-15 15:05:36
@article{c67e4246-3ec7-4d1a-aac0-c11a2f1f0116,
  abstract     = {This work presents a comparison of the crystal packing of three eukaryotic membrane proteins: human aquaporin 1, human aquaporin 5 and a spinach plasma membrane aquaporin. All were purified from expression constructs both with and without affinity tags. With the exception of tagged aquaporin 1, all constructs yielded crystals. Two significant effects of the affinity tags were observed: crystals containing a tag typically diffracted to lower resolution than those from constructs encoding the protein sequence alone and constructs without a tag frequently produced crystals that suffered from merohedral twinning. Twinning is a challenging crystallographic problem that can seriously hinder solution of the structure. Thus, for integral membrane proteins, the addition of an affinity tag may help to disrupt the approximate symmetry of the protein and thereby reduce or avoid merohedral twinning.},
  author       = {Backmark, Anna and Nyblom, Maria and Tornroth-Horsefield, Susanna and Kosinska-Eriksson, Urszula and Nordén, Kristina and Agemark, Maria and Kjellbom, Per and Johanson, Urban and Hedfalk, Kristina and Lindkvist- ersson, Karin and Neutze, Richard and Horsefield, Rob},
  issn         = {1399-0047},
  language     = {eng},
  pages        = {1183--1186},
  publisher    = {International Union of Crystallography},
  series       = {Acta Crystallographica. Section D: Biological Crystallography},
  title        = {Affinity tags can reduce merohedral twinning of membrane protein crystals},
  url          = {http://dx.doi.org/10.1107/S090744490802948X},
  volume       = {64},
  year         = {2008},
}