Synthesis of TOAC spin-labeled proteins and reconstitution in lipid membranes

Christine B. Karim, Zhiwen Zhang, David D. Thomas

Research output: Contribution to journalArticlepeer-review

38 Scopus citations


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 α-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.

Original languageEnglish (US)
Pages (from-to)42-49
Number of pages8
JournalNature Protocols
Issue number1
StatePublished - Feb 2007

Bibliographical note

Funding Information:
ACKNOWLEDGMENTS The authors thank Henriette Remmer at the Department of Biological Chemistry, University of Michigan, for advice about experimental details. This work was supported by a grant from the National Institutes of Health to D.D.T. (GM27906).


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