Journal
NATURE PROTOCOLS
Volume 2, Issue 10, Pages 2332-2338Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2007.329
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Funding
- NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING [R01EB005161, R01EB001966, P41EB002031] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [F32GM065833, R01GM075877] Funding Source: NIH RePORTER
- NIBIB NIH HHS [R01EB005161, P41EB002031, R01EB 001966] Funding Source: Medline
- NIGMS NIH HHS [F32GM65833, R01GM075877] Funding Source: Medline
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Magnetically aligned bicelles are an excellent medium for structure determination of isotopically labeled membrane proteins by solid-state NMR spectroscopy. Bicelles are a mixture of long- and short-chain phospholipids that form bilayers in an aqueous medium and align spontaneously in a high magnetic field, for example that of an NMR spectrometer with a H-1 resonance frequency between 400 and 900 MHz. Importantly, membrane proteins have been shown to be fully functional in these fully hydrated, planar bilayers under physiological conditions of pH and temperature. We describe a protocol for preparing stable protein-containing bicelles samples that yield high-resolution solid-state NMR spectra. Depending on the details of the protein and its behavior in the lipids, the time for sample preparation can vary from a few hours to several days.
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