Abstract
Metabolic labeling of proteins with the methionine surrogate azidonorleucine can be targeted exclusively to specified cells through expression of a mutant methionyl-tRNA synthetase (MetRS). In complex cellular mixtures, proteins made in cells that express the mutant synthetase can be tagged with affinity reagents (for detection or enrichment) or fluorescent dyes (for imaging). Proteins made in cells that do not express the mutant synthetase are neither labeled nor detected.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Affinity Labels / metabolism*
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Alanine / analogs & derivatives
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Alanine / metabolism
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Amino Acyl-tRNA Synthetases / genetics
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Amino Acyl-tRNA Synthetases / metabolism*
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Animals
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Cell Line
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Coculture Techniques
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Macrophages, Alveolar / metabolism
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Macrophages, Alveolar / microbiology
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Methionine-tRNA Ligase / genetics
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Methionine-tRNA Ligase / metabolism*
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Mice
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Mutagenesis, Site-Directed
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Mutation
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Norleucine / analogs & derivatives
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Norleucine / metabolism
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Protein Biosynthesis
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Proteins / metabolism*
Substances
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Affinity Labels
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Proteins
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Norleucine
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Amino Acyl-tRNA Synthetases
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Methionine-tRNA Ligase
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Alanine