In order to develop triple-resonance solid-state NMR spectroscopy of membrane proteins, we have implemented several different (13)C labeling schemes with the purpose of overcoming the interfering effects of (13)C-(13)C dipole-dipole couplings in stationary samples. The membrane-bound form of the major coat protein of the filamentous bacteriophage Pf1 was used as an example of a well-characterized helical membrane protein. Aligned protein samples randomly enriched to 35\% (13)C in all sites and metabolically labeled from bacterial growth on media containing 2-(13)C-glycerol or 1,3-(13)C-glycerol enables direct (13)C detection in solid-state NMR experiments without the need for homonuclear (13)C-(13)C dipole-dipole decoupling. The (13)C-detected NMR spectra of Pf1 coat protein show a substantial increase in sensitivity compared to the equivalent (15)N-detected spectra. The isotopic labeling pattern was analyzed for 2-(13)C-glycerol and 1,3-(13)C-glycerol as metabolic precursors by solution-state NMR of micelle samples. Polarization inversion spin exchange at the magic angle (PISEMA) and other solid-state NMR experiments work well on 35\% random fractionally and metabolically tailored (13)C-labeled samples, in contrast to their failure with conventional 100\% uniformly (13)C-labeled samples.
%0 Journal Article
%1 sinha_tailoring_2007
%A Sinha, Neeraj
%A Filipp, Fabian V
%A Jairam, Lena
%A Park, Sang Ho
%A Bradley, Joel
%A Opella, Stanley J
%D 2007
%J Magn Reson Chem
%K Bacteriophages,Carbon Core Isotopes,Magnetic Proteins Proteins,Viral Resonance Spectroscopy,Membrane Structural
%P S107----115
%R 10.1002/mrc.2121
%T Tailoring 13C labeling for triple-resonance solid-state \NMR\ experiments on aligned samples of proteins
%V 45 Suppl 1
%X In order to develop triple-resonance solid-state NMR spectroscopy of membrane proteins, we have implemented several different (13)C labeling schemes with the purpose of overcoming the interfering effects of (13)C-(13)C dipole-dipole couplings in stationary samples. The membrane-bound form of the major coat protein of the filamentous bacteriophage Pf1 was used as an example of a well-characterized helical membrane protein. Aligned protein samples randomly enriched to 35\% (13)C in all sites and metabolically labeled from bacterial growth on media containing 2-(13)C-glycerol or 1,3-(13)C-glycerol enables direct (13)C detection in solid-state NMR experiments without the need for homonuclear (13)C-(13)C dipole-dipole decoupling. The (13)C-detected NMR spectra of Pf1 coat protein show a substantial increase in sensitivity compared to the equivalent (15)N-detected spectra. The isotopic labeling pattern was analyzed for 2-(13)C-glycerol and 1,3-(13)C-glycerol as metabolic precursors by solution-state NMR of micelle samples. Polarization inversion spin exchange at the magic angle (PISEMA) and other solid-state NMR experiments work well on 35\% random fractionally and metabolically tailored (13)C-labeled samples, in contrast to their failure with conventional 100\% uniformly (13)C-labeled samples.
@article{sinha_tailoring_2007,
abstract = {In order to develop triple-resonance solid-state NMR spectroscopy of membrane proteins, we have implemented several different (13)C labeling schemes with the purpose of overcoming the interfering effects of (13)C-(13)C dipole-dipole couplings in stationary samples. The membrane-bound form of the major coat protein of the filamentous bacteriophage Pf1 was used as an example of a well-characterized helical membrane protein. Aligned protein samples randomly enriched to 35{\%} (13)C in all sites and metabolically labeled from bacterial growth on media containing [2-(13)C]-glycerol or [1,3-(13)C]-glycerol enables direct (13)C detection in solid-state NMR experiments without the need for homonuclear (13)C-(13)C dipole-dipole decoupling. The (13)C-detected NMR spectra of Pf1 coat protein show a substantial increase in sensitivity compared to the equivalent (15)N-detected spectra. The isotopic labeling pattern was analyzed for [2-(13)C]-glycerol and [1,3-(13)C]-glycerol as metabolic precursors by solution-state NMR of micelle samples. Polarization inversion spin exchange at the magic angle (PISEMA) and other solid-state NMR experiments work well on 35{\%} random fractionally and metabolically tailored (13)C-labeled samples, in contrast to their failure with conventional 100{\%} uniformly (13)C-labeled samples.},
added-at = {2017-03-14T02:48:56.000+0100},
author = {Sinha, Neeraj and Filipp, Fabian V and Jairam, Lena and Park, Sang Ho and Bradley, Joel and Opella, Stanley J},
biburl = {https://www.bibsonomy.org/bibtex/2f1f7a06a44e81caaa71d7f6fa6b231d3/nmrresource},
doi = {10.1002/mrc.2121},
interhash = {80f7f463ed16f43a7ebaf0970093220e},
intrahash = {f1f7a06a44e81caaa71d7f6fa6b231d3},
issn = {1097-458X},
journal = {Magn Reson Chem},
keywords = {Bacteriophages,Carbon Core Isotopes,Magnetic Proteins Proteins,Viral Resonance Spectroscopy,Membrane Structural},
month = dec,
pages = {S107----115},
pmid = {18157808},
timestamp = {2017-03-14T02:49:21.000+0100},
title = {{Tailoring 13C labeling for triple-resonance solid-state {\{}NMR{\}} experiments on aligned samples of proteins}},
volume = {45 Suppl 1},
year = 2007
}