Publication date: Available online 2 December 2016 Source:Journal of Magnetic Resonance
Author(s): Svetlana Kucher, Sergei Korneev, Swati Tyagi, Ronja Apfelbaum, Dina Grohmann, Edward A. Lemke, Johann P. Klare, Heinz-Jürgen Steinhoff, Daniel Klose
Site-directed spin labeling for EPR- and NMR spectroscopy has mainly been achieved exploiting the specific reactivity of cysteines. For proteins with native cysteines or for in vivo applications, an alternative coupling strategy is required. In these cases click chemistry offers major benefits by providing a fast and highly selective, biocompatible reaction between azide and alkyne groups. Here, we establish click chemistry as a tool to target unnatural amino acids in vitro and in vivo using azide- and alkyne-functionalized spin labels. The approach is compatible with a variety of labels including reduction-sensitive nitroxides. Comparing spin labeling efficiencies from the copper-free with the strongly reducing copper(I)-catalyzed azide-alkyne click reaction, we find that the faster kinetics for the catalyzed reaction outrun reduction of the labile nitroxide spin labels and allow quantitative labeling yields within short reaction times. Inter-spin distance measurements demonstrate that the novel side chain is suitable for paramagnetic NMR- or EPR-based conformational studies of macromolecular complexes. Graphical abstract
Gd(iii) and Mn(ii) complexes for dynamic nuclear polarization: small molecular chelate polarizing agents and applications with site-directed spin labeling of proteins #DNPNMR
From The DNP-NMR Blog:
Gd(iii) and Mn(ii) complexes for dynamic nuclear polarization: small molecular chelate polarizing agents and applications with site-directed spin labeling of proteins #DNPNMR
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Kaushik, M., et al., Gd(iii) and Mn(ii) complexes for dynamic nuclear polarization: small molecular chelate polarizing agents and applications with site-directed spin labeling of proteins. Phys Chem Chem Phys, 2016. 18(39): p. 27205-27218.
https://www.ncbi.nlm.nih.gov/pubmed/27545112
[NMR paper] In vivo and in vitro metabolism of a novel ?2-adrenoceptor agonist, trantinterol: metabolites isolation and identification by LC-MS/MS and NMR.
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Abstract
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Introduction: Nuclear magnetic resonance (NMR) applications in drug discovery are classified into two categories: ligand-based methods and protein-based methods. The latter is based on the observation of the (1)H-(15)N HSQC spectra of a protein with and without lead compounds. However, in order to take this...
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Shanks, Jacqueline Vanni (1989) Metabolic engineering applications of in vivo P and C NMR studies of Saccharomyces cerevisiae. Dissertation (Ph.D.), California Institute of Technology. http://resolver.caltech.edu/CaltechETD:etd-01242007-080430
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Magn Reson Med. 1993 Feb;29(2):252-5
Authors: Vion-Dury J, Machy P, Confort-Gouny S, Leserman L, Cozzone PJ
Liposomes containing dexamethasone phosphate (DMp) were covalently coupled to protein A and then incubated with murine L929 fibroblast and RDM4...