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Crystallization of Human Aquaglyceroporins for Neutron Diffraction Studies

de Maré, Sofia W. LU ; Hyld Steffen, Jonas ; Missel, Julie W. ; Gerdt Laursen, Amalie ; Gotfryd, Kamil ; Fisher, Zoë LU ; Amiry-Moghaddam, Mahmood Reza ; Gourdon, Pontus LU and Lindkvist-Petersson, Karin LU (2025) In ACS Omega 10(8). p.7806-7812
Abstract

Aquaglyceroporins are channels that facilitate the flux of glycerol and water across lipid bilayers. Although structural information is available for several aquaglyceroporins, the details of how water and glycerol selectivity are maintained and how protons are excluded remain elusive. An approach to obtaining data on the hydrogen atom positions is to apply neutron macromolecular crystallography. Here, we present strategies to obtain large crystals suitable for neutron diffraction experiments by assessing a range of different methods, including new procedures for protein purification and crystallization. By applying long incubation times, macroseeding, and/or optimization of detergents, millimeter-sized crystals with different... (More)

Aquaglyceroporins are channels that facilitate the flux of glycerol and water across lipid bilayers. Although structural information is available for several aquaglyceroporins, the details of how water and glycerol selectivity are maintained and how protons are excluded remain elusive. An approach to obtaining data on the hydrogen atom positions is to apply neutron macromolecular crystallography. Here, we present strategies to obtain large crystals suitable for neutron diffraction experiments by assessing a range of different methods, including new procedures for protein purification and crystallization. By applying long incubation times, macroseeding, and/or optimization of detergents, millimeter-sized crystals with different morphologies were obtained, and their diffraction quality was assessed by exposure to X-rays. The data presented here lay the foundation for continued crystallization efforts targeting aquaporins and other membrane proteins for neutron diffraction experiments.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
ACS Omega
volume
10
issue
8
pages
7 pages
publisher
The American Chemical Society (ACS)
external identifiers
  • pmid:40060823
  • scopus:86000382525
ISSN
2470-1343
DOI
10.1021/acsomega.4c08284
language
English
LU publication?
yes
id
e890f6ac-ade7-4bb6-b78b-7771fdee2331
date added to LUP
2025-06-23 10:46:53
date last changed
2025-07-07 11:06:29
@article{e890f6ac-ade7-4bb6-b78b-7771fdee2331,
  abstract     = {{<p>Aquaglyceroporins are channels that facilitate the flux of glycerol and water across lipid bilayers. Although structural information is available for several aquaglyceroporins, the details of how water and glycerol selectivity are maintained and how protons are excluded remain elusive. An approach to obtaining data on the hydrogen atom positions is to apply neutron macromolecular crystallography. Here, we present strategies to obtain large crystals suitable for neutron diffraction experiments by assessing a range of different methods, including new procedures for protein purification and crystallization. By applying long incubation times, macroseeding, and/or optimization of detergents, millimeter-sized crystals with different morphologies were obtained, and their diffraction quality was assessed by exposure to X-rays. The data presented here lay the foundation for continued crystallization efforts targeting aquaporins and other membrane proteins for neutron diffraction experiments.</p>}},
  author       = {{de Maré, Sofia W. and Hyld Steffen, Jonas and Missel, Julie W. and Gerdt Laursen, Amalie and Gotfryd, Kamil and Fisher, Zoë and Amiry-Moghaddam, Mahmood Reza and Gourdon, Pontus and Lindkvist-Petersson, Karin}},
  issn         = {{2470-1343}},
  language     = {{eng}},
  number       = {{8}},
  pages        = {{7806--7812}},
  publisher    = {{The American Chemical Society (ACS)}},
  series       = {{ACS Omega}},
  title        = {{Crystallization of Human Aquaglyceroporins for Neutron Diffraction Studies}},
  url          = {{http://dx.doi.org/10.1021/acsomega.4c08284}},
  doi          = {{10.1021/acsomega.4c08284}},
  volume       = {{10}},
  year         = {{2025}},
}