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

Christine B Karim, Zhiwen Zhang, David D Thomas

Research output: Contribution to journalArticle

34 Scopus citations

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 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
Volume2
Issue number1
DOIs
StatePublished - Feb 1 2007

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