Protocol abstract
Nature Protocols 2, - 2332 - 2338 (2007)
Published online: 20 September 2007 | doi:10.1038/nprot.2007.329
Subject Categories: Spectroscopy and structural analysis | Biochemistry and protein analysis
Bicelle samples for solid-state NMR of membrane proteins
Anna A De Angelis1 & Stanley J Opella1
Abstract
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 1H 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.
- Department of Chemistry and Biochemistry, University of California at San Diego, 9500 Gilman Drive, La Jolla, California 92037, USA.
Correspondence to: Stanley J Opella1 e-mail: sopella@ucsd.edu
nature-products
A-Z product listing
- Bath sonicator(Branson)
- Benchtop centrifuge(Eppendorff)
- Flat-bottomed NMR tubes(New Era Enterprises)
- Glass syringes(Hamilton)
- Heat block(Fisher Scientific)
- Long-chain lipid(s)(Avanti Polar Lipids)
