Abstract
Sample solubility is essential for structural studies of proteins by solution NMR. Attachment of a solubility enhancement tag, such as GB1, MBP and thioredoxin, to a target protein has been used for this purpose. However, signal overlap of the tag with the target protein often made the spectral analysis difficult. Here we report a sortase-mediated protein ligation method to eliminate NMR signals arising from the tag by preparing the isotopically labeled target protein attached with the non-labeled GB1 tag at the C-terminus.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aminoacyltransferases / metabolism
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Bacterial Proteins / metabolism
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Cysteine Endopeptidases / metabolism
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Nuclear Magnetic Resonance, Biomolecular*
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Protein Conformation
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Proteins / chemistry*
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Proteins / genetics
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Proteins / metabolism*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Solubility
Substances
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Bacterial Proteins
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Proteins
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Recombinant Fusion Proteins
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Aminoacyltransferases
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sortase A
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Cysteine Endopeptidases