| Solid-state NMR analysis of membrane proteins and protein aggregates by proton detected spectroscopy. | |
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MedLine Citation:
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PMID: 22986689 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Solid-state NMR has emerged as an important tool for structural biology and chemistry, capable of solving atomic-resolution structures for proteins in membrane-bound and aggregated states. Proton detection methods have been recently realized under fast magic-angle spinning conditions, providing large sensitivity enhancements for efficient examination of uniformly labeled proteins. The first and often most challenging step of protein structure determination by NMR is the site-specific resonance assignment. Here we demonstrate resonance assignments based on high-sensitivity proton-detected three-dimensional experiments for samples of different physical states, including a fully-protonated small protein (GB1, 6 kDa), a deuterated microcrystalline protein (DsbA, 21 kDa), a membrane protein (DsbB, 20 kDa) prepared in a lipid environment, and the extended core of a fibrillar protein (α-synuclein, 14 kDa). In our implementation of these experiments, including CONH, CO(CA)NH, CANH, CA(CO)NH, CBCANH, and CBCA(CO)NH, dipolar-based polarization transfer methods have been chosen for optimal efficiency for relatively high protonation levels (full protonation or 100 % amide proton), fast magic-angle spinning conditions (40 kHz) and moderate proton decoupling power levels. Each H-N pair correlates exclusively to either intra- or inter-residue carbons, but not both, to maximize spectral resolution. Experiment time can be reduced by at least a factor of 10 by using proton detection in comparison to carbon detection. These high-sensitivity experiments are especially important for membrane proteins, which often have rather low expression yield. Proton-detection based experiments are expected to play an important role in accelerating protein structure elucidation by solid-state NMR with the improved sensitivity and resolution. |
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Authors:
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Donghua H Zhou; Andrew J Nieuwkoop; Deborah A Berthold; Gemma Comellas; Lindsay J Sperling; Ming Tang; Gautam J Shah; Elliott J Brea; Luisel R Lemkau; Chad M Rienstra |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2012-9-18 |
Journal Detail:
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Title: Journal of biomolecular NMR Volume: - ISSN: 1573-5001 ISO Abbreviation: J. Biomol. NMR Publication Date: 2012 Sep |
Date Detail:
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Created Date: 2012-9-18 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9110829 Medline TA: J Biomol NMR Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Affiliation:
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Department of Physics, Oklahoma State University, Stillwater, OK, 74074, USA, donghua@okstate.edu. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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