Macroscopic orientation of natural and model membranes for structural studies.
One approach for obtaining high-resolution structural and functional information for biomembranes and their proteins is by static solid-state NMR of oriented systems. Here, a general procedure to align fully functional biological membranes containing large membrane proteins (Mr >30,000) is de...
Main Authors: | , , , , , , , |
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Format: | Journal article |
Language: | English |
Published: |
1997
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_version_ | 1797079724040650752 |
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author | Gröbner, G Taylor, A Williamson, P Choi, G Glaubitz, C Watts, J de Grip, W Watts, A |
author_facet | Gröbner, G Taylor, A Williamson, P Choi, G Glaubitz, C Watts, J de Grip, W Watts, A |
author_sort | Gröbner, G |
collection | OXFORD |
description | One approach for obtaining high-resolution structural and functional information for biomembranes and their proteins is by static solid-state NMR of oriented systems. Here, a general procedure to align fully functional biological membranes containing large membrane proteins (Mr >30,000) is described. The method, based on the isopotential spin-dry ultracentrifugation technique, relies on the centrifugation of membrane fragments onto a support with simultaneous, or subsequent, partial evaporation of the solvent which aids alignment. The quality of orientation, as shown by the mosaic spread of the samples, was monitored by static solid-state 31P NMR for the phospholipids and by 2H NMR for a deuterated retinal in bovine rhodopsin. The generality of this method is demonstrated with three different membranes containing bovine rhodopsin in reconstituted bilayers, natural membranes with the red cell anion exchange transport protein in erythrocytes, band 3, and the nicotinic acetylcholine receptor. |
first_indexed | 2024-03-07T00:49:49Z |
format | Journal article |
id | oxford-uuid:85fd401c-13c1-4233-b7b6-1c04051160ce |
institution | University of Oxford |
language | English |
last_indexed | 2024-03-07T00:49:49Z |
publishDate | 1997 |
record_format | dspace |
spelling | oxford-uuid:85fd401c-13c1-4233-b7b6-1c04051160ce2022-03-26T22:01:12ZMacroscopic orientation of natural and model membranes for structural studies.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:85fd401c-13c1-4233-b7b6-1c04051160ceEnglishSymplectic Elements at Oxford1997Gröbner, GTaylor, AWilliamson, PChoi, GGlaubitz, CWatts, Jde Grip, WWatts, AOne approach for obtaining high-resolution structural and functional information for biomembranes and their proteins is by static solid-state NMR of oriented systems. Here, a general procedure to align fully functional biological membranes containing large membrane proteins (Mr >30,000) is described. The method, based on the isopotential spin-dry ultracentrifugation technique, relies on the centrifugation of membrane fragments onto a support with simultaneous, or subsequent, partial evaporation of the solvent which aids alignment. The quality of orientation, as shown by the mosaic spread of the samples, was monitored by static solid-state 31P NMR for the phospholipids and by 2H NMR for a deuterated retinal in bovine rhodopsin. The generality of this method is demonstrated with three different membranes containing bovine rhodopsin in reconstituted bilayers, natural membranes with the red cell anion exchange transport protein in erythrocytes, band 3, and the nicotinic acetylcholine receptor. |
spellingShingle | Gröbner, G Taylor, A Williamson, P Choi, G Glaubitz, C Watts, J de Grip, W Watts, A Macroscopic orientation of natural and model membranes for structural studies. |
title | Macroscopic orientation of natural and model membranes for structural studies. |
title_full | Macroscopic orientation of natural and model membranes for structural studies. |
title_fullStr | Macroscopic orientation of natural and model membranes for structural studies. |
title_full_unstemmed | Macroscopic orientation of natural and model membranes for structural studies. |
title_short | Macroscopic orientation of natural and model membranes for structural studies. |
title_sort | macroscopic orientation of natural and model membranes for structural studies |
work_keys_str_mv | AT grobnerg macroscopicorientationofnaturalandmodelmembranesforstructuralstudies AT taylora macroscopicorientationofnaturalandmodelmembranesforstructuralstudies AT williamsonp macroscopicorientationofnaturalandmodelmembranesforstructuralstudies AT choig macroscopicorientationofnaturalandmodelmembranesforstructuralstudies AT glaubitzc macroscopicorientationofnaturalandmodelmembranesforstructuralstudies AT wattsj macroscopicorientationofnaturalandmodelmembranesforstructuralstudies AT degripw macroscopicorientationofnaturalandmodelmembranesforstructuralstudies AT wattsa macroscopicorientationofnaturalandmodelmembranesforstructuralstudies |