Abstract
Site-specific protein cleavage is essential for many protein-production protocols and typically requires proteases. We report the development of a chemical protein-cleavage method that is achieved through the use of a sequence-specific nickel-assisted cleavage (SNAC)-tag. We demonstrate that the SNAC-tag can be inserted before both water-soluble and membrane proteins to achieve fusion protein cleavage under biocompatible conditions with efficiency comparable to that of enzymes, and that the method works even when enzymatic cleavages fail.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Biocompatible Materials
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Chromatography, High Pressure Liquid
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Computational Biology
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DNA / chemistry
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Endopeptidases / genetics
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Endopeptidases / metabolism
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Enzymes / chemistry*
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Escherichia coli / metabolism
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Genetic Techniques
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Hydrolysis
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Mass Spectrometry
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Nickel / chemistry*
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Peptide Library
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Peptides / chemistry
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Protein Domains
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Proteins / chemistry*
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Proteolysis
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Recombinant Proteins / chemistry
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Substrate Specificity
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Temperature
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Thrombin / chemistry
Substances
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Biocompatible Materials
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Enzymes
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Peptide Library
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Peptides
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Proteins
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Recombinant Proteins
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Nickel
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DNA
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Endopeptidases
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TEV protease
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Thrombin