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Prospects for membrane protein crystals in NMX

Hjorth-Jensen, Samuel John LU ; Oksanen, Esko LU ; Nissen, Poul and Sørensen, Thomas Lykke Møller (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.

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Please use this url to cite or link to this publication:
author
; ; and
organization
publishing date
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}},
}