Crystallization of Human Aquaglyceroporins for Neutron Diffraction Studies
(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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- author
- 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
- organization
- publishing date
- 2025-03
- 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}}, }