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Fractional deuteration applied to biomolecular solid-state NMR spectroscopy.

ABSTRACT: Solid-state Nuclear Magnetic Resonance can provide detailed insight into structural and dynamical aspects of complex biomolecules. With increasing molecular size, advanced approaches for spectral simplification and the detection of medium to long-range contacts become of critical relevance. We have analyzed the protonation pattern of a membrane-embedded ion channel that was obtained from bacterial expression using protonated precursors and D(2)O medium. We find an overall reduction of 50% in protein protonation. High levels of deuteration at H(?) and H(?) positions reduce spectral congestion in ((1)H,(13)C,(15)N) correlation experiments and generate a transfer profile in longitudinal mixing schemes that can be tuned to specific resonance frequencies. At the same time, residual protons are predominantly found at amino-acid side-chain positions enhancing the prospects for obtaining side-chain resonance assignments and for detecting medium to long-range contacts. Fractional deuteration thus provides a powerful means to aid the structural analysis of complex biomolecules by solid-state NMR.

SUBMITTER: Nand D

PROVIDER: S-EPMC3277825 | biostudies-literature | 2012 Feb

REPOSITORIES: biostudies-literature

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Fractional deuteration applied to biomolecular solid-state NMR spectroscopy.

Nand Deepak D Cukkemane Abhishek A Becker Stefan S Baldus Marc M

Journal of biomolecular NMR 20111122 2

Solid-state Nuclear Magnetic Resonance can provide detailed insight into structural and dynamical aspects of complex biomolecules. With increasing molecular size, advanced approaches for spectral simplification and the detection of medium to long-range contacts become of critical relevance. We have analyzed the protonation pattern of a membrane-embedded ion channel that was obtained from bacterial expression using protonated precursors and D(2)O medium. We find an overall reduction of 50% in pro ...[more]

PMID: 22105305

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Fractional deuteration applied to biomolecular solid-state NMR spectroscopy.

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Fractional deuteration applied to biomolecular solid-state NMR spectroscopy.

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