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. 2011 Jan;20(1):224-8.
doi: 10.1002/pro.545.

Site-specific ¹⁹F NMR chemical shift and side chain relaxation analysis of a membrane protein labeled with an unnatural amino acid

Pan Shi  1 , Hu Wang, Zhaoyong Xi, Chaowei Shi, Ying Xiong, Changlin Tian
Affiliations

Site-specific ¹⁹F NMR chemical shift and side chain relaxation analysis of a membrane protein labeled with an unnatural amino acid

Pan Shi et al. Protein Sci. 2011 Jan.

Abstract

Site-specific ¹⁹F chemical shift and side chain relaxation analysis can be applied on large size proteins. Here, one-dimensional ¹⁹F spectra and T₁, T₂ relaxation data were acquired on a SH3 domain in aqueous buffer containing 60% glycerol, and a nine-transmembrane helices membrane protein diacyl-glycerol kinase (DAGK) in dodecyl phosphochoine (DPC) micelles. The high quality of the data indicates that this method can be applied to site-specifically analyze side chain internal mobility of membrane proteins or large size proteins.

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Figures

Figure 1
Figure 1
19F chemical shift and T1, T2 relaxation analysis of the Phe7-tfmF-SH3 domain in aqueous buffer. Different 19F chemical shifts (A, D) and T1 relaxation (B, E), T2 relaxation values (C, F) are observed in the absence (A, B, C) or presence (D, E, F) of a peptide ligand P868.
Figure 2
Figure 2
Diffusion coefficient measurements of water (A), 19F chemical shift (B) and T1 relaxation (C) and T2 relaxation analysis (D) from the sample of Phe7-tfmF-SH3 in aqueous buffer containing 60% glycerol.
Figure 3
Figure 3
One-dimensional 19F chemical shift (A), T1 relaxation (B) and T2 relaxation analysis (C) of a membrane protein DAGK labelled at position 31 with tfmF in DPC micelles (Phe31-tfmF-DAGK/DPC).
See this image and copyright information in PMC

References

    1. Lipchock JM, Loria JP. Monitoring molecular interactions by NMR. Methods Mol Biol. 2009;490:115–134. - PubMed
    1. Haggie PM, Brindle KM. Mitochondrial citrate synthase is immobilized in vivo. J Biol Chem. 1999;274:3941–3945. - PubMed
    1. Williams SP, Haggie PM, Brindle KM. 19F NMR measurements of the rotational mobility of proteins in vivo. Biophys J. 1997;72:490–498. - PMC - PubMed
    1. Jackson JC, Hammill JT, Mehl RA. Site-specific incorporation of a 19F-amino acid into proteins as an NMR probe for characterizing protein structure and reactivity. J Am Chem Soc. 2007;129:1160–1166. - PubMed
    1. Hammill JT, Miyake-Stoner S, Hazen JL, Jackson JC, Mehl RA. Preparation of site-specifically labeled fluorinated proteins for 19F-NMR structural characterization. Nat Protoc. 2007;2:2601–2607. - PubMed

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