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

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

doi:10.1038/nprot.2007.2

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

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

Chemical and Structural Biology (TMED)

Research output: Contribution to journal › Article › peer-review

36 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 language	English (US)
Pages (from-to)	42-49
Number of pages	8
Journal	Nature Protocols
Volume	2
Issue number	1
DOIs	https://doi.org/10.1038/nprot.2007.2
State	Published - 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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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.",

author = "Karim, {Christine B.} and Zhiwen Zhang and Thomas, {David D.}",

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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AU - Thomas, David D.

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

Abstract

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