Prospects for membrane protein crystals in NMX
(2020) In Methods in Enzymology 634. p.47-68- Abstract
Adding hydrogen atoms and protonation states to structures of membrane proteins requires successful implementation of neutron macromolecular crystallography (NMX). This information would significantly increase our fundamental understanding of the transport processes membrane proteins undertake. To grow the large crystals needed for NMX studies requires significant amounts of stable protein, but once that challenge is overcome there is no intrinsic property of membrane proteins preventing the growth of large crystals per se. The calcium-transporting P-type ATPase (SERCA) has been thoroughly characterized biochemically and structurally over decades. We have extended our crystallization efforts to assess the feasibility of growing SERCA... (More)
Adding hydrogen atoms and protonation states to structures of membrane proteins requires successful implementation of neutron macromolecular crystallography (NMX). This information would significantly increase our fundamental understanding of the transport processes membrane proteins undertake. To grow the large crystals needed for NMX studies requires significant amounts of stable protein, but once that challenge is overcome there is no intrinsic property of membrane proteins preventing the growth of large crystals per se. The calcium-transporting P-type ATPase (SERCA) has been thoroughly characterized biochemically and structurally over decades. We have extended our crystallization efforts to assess the feasibility of growing SERCA crystals for NMX—exploring microdialysis and capillary counterdiffusion crystallization techniques as alternatives to the traditional vapor diffusion crystallization experiment. Both methods possess crystallization dynamics favorable for maximizing crystal size and we used them to facilitate the growth of large crystals, validating these approaches for membrane protein crystallization for NMX.
(Less)
- author
- Hjorth-Jensen, Samuel John LU ; Oksanen, Esko LU ; Nissen, Poul and Sørensen, Thomas Lykke Møller
- organization
- publishing date
- 2020-01-10
- type
- Chapter in Book/Report/Conference proceeding
- publication status
- published
- subject
- keywords
- Crystallization techniques, Membrane protein structure, MX, Neutron diffraction, NMX, Protein crystallography, Structural biology
- host publication
- Neutron Crystallography in Structural Biology
- series title
- Methods in Enzymology
- editor
- Moody, Peter C.E.
- volume
- 634
- pages
- 22 pages
- publisher
- Academic Press
- external identifiers
-
- scopus:85077699655
- pmid:32093842
- ISSN
- 0076-6879
- 1557-7988
- DOI
- 10.1016/bs.mie.2019.11.019
- language
- English
- LU publication?
- yes
- id
- 0a2f2db8-412c-4b39-b138-fafcd2b6bbcc
- date added to LUP
- 2020-01-29 08:48:28
- date last changed
- 2024-04-03 02:00:00
@inbook{0a2f2db8-412c-4b39-b138-fafcd2b6bbcc, abstract = {{<p>Adding hydrogen atoms and protonation states to structures of membrane proteins requires successful implementation of neutron macromolecular crystallography (NMX). This information would significantly increase our fundamental understanding of the transport processes membrane proteins undertake. To grow the large crystals needed for NMX studies requires significant amounts of stable protein, but once that challenge is overcome there is no intrinsic property of membrane proteins preventing the growth of large crystals per se. The calcium-transporting P-type ATPase (SERCA) has been thoroughly characterized biochemically and structurally over decades. We have extended our crystallization efforts to assess the feasibility of growing SERCA crystals for NMX—exploring microdialysis and capillary counterdiffusion crystallization techniques as alternatives to the traditional vapor diffusion crystallization experiment. Both methods possess crystallization dynamics favorable for maximizing crystal size and we used them to facilitate the growth of large crystals, validating these approaches for membrane protein crystallization for NMX.</p>}}, author = {{Hjorth-Jensen, Samuel John and Oksanen, Esko and Nissen, Poul and Sørensen, Thomas Lykke Møller}}, booktitle = {{Neutron Crystallography in Structural Biology}}, editor = {{Moody, Peter C.E.}}, issn = {{0076-6879}}, keywords = {{Crystallization techniques; Membrane protein structure; MX; Neutron diffraction; NMX; Protein crystallography; Structural biology}}, language = {{eng}}, month = {{01}}, pages = {{47--68}}, publisher = {{Academic Press}}, series = {{Methods in Enzymology}}, title = {{Prospects for membrane protein crystals in NMX}}, url = {{http://dx.doi.org/10.1016/bs.mie.2019.11.019}}, doi = {{10.1016/bs.mie.2019.11.019}}, volume = {{634}}, year = {{2020}}, }