Synthesis of TOAC spin-labeled proteins and reconstitution in lipid membranes |
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| Authors: | Karim Christine B Zhang Zhiwen Thomas David D |
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| Institution: | Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA. cbk@ddt.biochem.umn.edu |
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| Abstract: | A procedure is described for the synthetic incorporation into membrane proteins of the non-natural amino acid TOAC (2,2,6,6-tetramethyl-piperidine-1-oxyl-4-amino-4-carboxylic acid), which is coupled rigidly to the alpha-carbon, providing direct detection of peptide backbone dynamics by electron paramagnetic resonance (EPR). Also included is a protocol for the functional reconstitution of the spin-labeled protein in lipid vesicles. This protocol can be completed in 17 d. |
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