| Protocols for the sequential solid-state NMR spectroscopic assignment of a uniformly labeled 25 kDa protein: HET-s(1-227). | |
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MedLine Citation:
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PMID: 20572250 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The sequence-specific resonance assignment of a protein forms the basis for studies of molecular structure and dynamics, as well as to functional assay studies by NMR spectroscopy. Here we present a protocol for the sequential 13C and 15N resonance assignment of uniformly [15N,13C]-labeled proteins, based on a suite of complementary three-dimensional solid-state NMR spectroscopy experiments. It is directed towards the application to proteins with more than about 100 amino acid residues. The assignments rely on a walk along the backbone by using a combination of three experiments that correlate nitrogen and carbon spins, including the well-dispersed Cbeta resonances. Supplementary spectra that correlate further side-chain resonances can be important for identifying the amino acid type, and greatly assist the assignment process. We demonstrate the application of this assignment protocol for a crystalline preparation of the N-terminal globular domain of the HET-s prion, a 227-residue protein. |
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Authors:
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Anne Schuetz; Christian Wasmer; Birgit Habenstein; René Verel; Jason Greenwald; Roland Riek; Anja Böckmann; Beat H Meier |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Chembiochem : a European journal of chemical biology Volume: 11 ISSN: 1439-7633 ISO Abbreviation: Chembiochem Publication Date: 2010 Jul |
Date Detail:
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Created Date: 2010-07-26 Completed Date: 2010-11-02 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 100937360 Medline TA: Chembiochem Country: Germany |
Other Details:
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Languages: eng Pagination: 1543-51 Citation Subset: IM |
Affiliation:
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Physical Chemistry, ETH Zürich, 8093 Zürich, Switzerland. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Carbon Isotopes Magnetic Resonance Spectroscopy / methods* Nitrogen Isotopes Prions / chemistry Proteins / chemistry* Research Design |
| Chemical | |
Reg. No./Substance:
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0/Carbon Isotopes; 0/Nitrogen Isotopes; 0/Prions; 0/Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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