Kimata, Naoki and Pope, Andreyah and Rashid, Dawood and Reeves, Philip J and Smith, Steven O (2015) Sequential Structural Changes in Rhodopsin Occurring upon Photoactivation. In: Methods in Molecular Biology. Springer New York, pp. 159-171. ISBN 9781493923298. Official URL: https://doi.org/10.1007/978-1-4939-2330-4_11
Kimata, Naoki and Pope, Andreyah and Rashid, Dawood and Reeves, Philip J and Smith, Steven O (2015) Sequential Structural Changes in Rhodopsin Occurring upon Photoactivation. In: Methods in Molecular Biology. Springer New York, pp. 159-171. ISBN 9781493923298. Official URL: https://doi.org/10.1007/978-1-4939-2330-4_11
Kimata, Naoki and Pope, Andreyah and Rashid, Dawood and Reeves, Philip J and Smith, Steven O (2015) Sequential Structural Changes in Rhodopsin Occurring upon Photoactivation. In: Methods in Molecular Biology. Springer New York, pp. 159-171. ISBN 9781493923298. Official URL: https://doi.org/10.1007/978-1-4939-2330-4_11
Abstract
We describe the use of solid-state magic angle spinning NMR spectroscopy for characterizing the structure and dynamics of dark, inactive rhodopsin and the active metarhodopsin II intermediate. Solid-state NMR spectroscopy is well suited for structural measurements in both detergent micelles and membrane bilayer environments. We fi rst outline the methods for large-scale production of stable, functional rhodopsin containing 13 C- and 15 N-labeled amino acids. The expression methods make use of eukaryotic HEK293S cell lines that produce correctly folded, fully functional receptors. We subsequently describe the basic methods used for solid-state magic angle spinning NMR measurements of chemical shifts and dipolar couplings, which provide information on rhodopsin structure and dynamics, and describe the use of low-temperature methods to trap the active metarhodopsin II intermediate.
Item Type: | Book Section |
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Uncontrolled Keywords: | Cell Line; Humans; Lipid Bilayers; Rhodopsin; Amino Acids; Nuclear Magnetic Resonance, Biomolecular; Protein Structure, Secondary; Photochemistry |
Subjects: | Q Science > QH Natural history > QH301 Biology |
Divisions: | Faculty of Science and Health Faculty of Science and Health > Life Sciences, School of |
SWORD Depositor: | Unnamed user with email elements@essex.ac.uk |
Depositing User: | Unnamed user with email elements@essex.ac.uk |
Date Deposited: | 11 Dec 2016 10:49 |
Last Modified: | 05 Dec 2024 22:44 |
URI: | http://repository.essex.ac.uk/id/eprint/18468 |